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Jackson Immuno rabbit anti human grp75
$<t>GRP75</t> is an interactive partner of cholix. Polarized monolayers of Caco-2 cells were exposed apically to nontoxic full-length Chx (ntChx) anchored through a C-terminal hexa-histidine sequence to 100 nm diameter magnetic beads. After 15 min, cell membranes were isolated and separated into non-magnetic and magnetic fractions. a, 1-D SDS-PAGE gel of non-magnetic and magnetic captured membrane fractions. b, 2-D gel separation of proteins present in non-magnetic and magnetic capture membrane fractions with location of spot 16 identified by mass spectrometry as GRP75. c, Pull-down (PD) using ntChx probed with antibodies to either GRP75 or human growth hormone (hGH). d, Immunoprecipitation of Caco-2 cell lysate using ntChx probed with anti-GRP75 or control antibodies. e, 2-D Western blot of magnetic capture membrane fraction probed with anti-GRP75. f, ELISA-based, neutral pH, format where bound ntChx or the control protein bovine serum albumin (BSA) was used to capture GRP75, heat shock protein 60 (HSP60), glucose-regulated protein 78 (GRP78), or HSP90.$
Rabbit Anti Human Grp75, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 95/100, based on 374 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 95 stars, based on 374 article reviews

rabbit anti human grp75 - by Bioz Stars, 2026-09

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Article Title: GRP75 as a functional element of cholix transcytosis

Journal: Tissue Barriers

doi: 10.1080/21688370.2022.2039003

$GRP75 is an interactive partner of cholix. Polarized monolayers of Caco-2 cells were exposed apically to nontoxic full-length Chx (ntChx) anchored through a C-terminal hexa-histidine sequence to 100 nm diameter magnetic beads. After 15 min, cell membranes were isolated and separated into non-magnetic and magnetic fractions. a, 1-D SDS-PAGE gel of non-magnetic and magnetic captured membrane fractions. b, 2-D gel separation of proteins present in non-magnetic and magnetic capture membrane fractions with location of spot 16 identified by mass spectrometry as GRP75. c, Pull-down (PD) using ntChx probed with antibodies to either GRP75 or human growth hormone (hGH). d, Immunoprecipitation of Caco-2 cell lysate using ntChx probed with anti-GRP75 or control antibodies. e, 2-D Western blot of magnetic capture membrane fraction probed with anti-GRP75. f, ELISA-based, neutral pH, format where bound ntChx or the control protein bovine serum albumin (BSA) was used to capture GRP75, heat shock protein 60 (HSP60), glucose-regulated protein 78 (GRP78), or HSP90.$
Figure Legend Snippet: GRP75 is an interactive partner of cholix. Polarized monolayers of Caco-2 cells were exposed apically to nontoxic full-length Chx (ntChx) anchored through a C-terminal hexa-histidine sequence to 100 nm diameter magnetic beads. After 15 min, cell membranes were isolated and separated into non-magnetic and magnetic fractions. a, 1-D SDS-PAGE gel of non-magnetic and magnetic captured membrane fractions. b, 2-D gel separation of proteins present in non-magnetic and magnetic capture membrane fractions with location of spot 16 identified by mass spectrometry as GRP75. c, Pull-down (PD) using ntChx probed with antibodies to either GRP75 or human growth hormone (hGH). d, Immunoprecipitation of Caco-2 cell lysate using ntChx probed with anti-GRP75 or control antibodies. e, 2-D Western blot of magnetic capture membrane fraction probed with anti-GRP75. f, ELISA-based, neutral pH, format where bound ntChx or the control protein bovine serum albumin (BSA) was used to capture GRP75, heat shock protein 60 (HSP60), glucose-regulated protein 78 (GRP78), or HSP90.

Techniques Used: Sequencing, Magnetic Beads, Isolation, SDS Page, Membrane, Mass Spectrometry, Immunoprecipitation, Control, Western Blot, Enzyme-linked Immunosorbent Assay

GRP75 and perlecan are functionally involved in cholix transcytosis. Expression of three proteins identified by ntChx-magnetic bead capture were suppressed using a CRISPR/cas 9-mediated knockdown protocol that targeted either GRP75, HSPG2 , or KRT8 in Caco-2 cells. Confluent monolayers of the resulting cell lines having decreased levels of a, GRP75, b, perlecan, or c, cytokeratin 8 (CT8) were evaluated for their capacity for Chx-mediated transcytosis using Chx266-hGH and for the nonspecific movement of the control protein hGH over a 60 min time period. d, Transcytosis of Chx266-hGH across confluent monolayers of parent Caco-2 cells was evaluated following a simultaneous apical application of a monoclonal antibody against GRP75 or perlecan, with an isotype antibody recognizing interleukin (IL)-10 serving as a control. Densitometry measurements of individual Western blot bands are shown for graphical comparison.

Figure Legend Snippet: GRP75 and perlecan are functionally involved in cholix transcytosis. Expression of three proteins identified by ntChx-magnetic bead capture were suppressed using a CRISPR/cas 9-mediated knockdown protocol that targeted either GRP75, HSPG2 , or KRT8 in Caco-2 cells. Confluent monolayers of the resulting cell lines having decreased levels of a, GRP75, b, perlecan, or c, cytokeratin 8 (CT8) were evaluated for their capacity for Chx-mediated transcytosis using Chx266-hGH and for the nonspecific movement of the control protein hGH over a 60 min time period. d, Transcytosis of Chx266-hGH across confluent monolayers of parent Caco-2 cells was evaluated following a simultaneous apical application of a monoclonal antibody against GRP75 or perlecan, with an isotype antibody recognizing interleukin (IL)-10 serving as a control. Densitometry measurements of individual Western blot bands are shown for graphical comparison.

Techniques Used: Expressing, CRISPR, Knockdown, Control, Western Blot, Comparison

GRP75 associates with cholix in apical endosomes. Immunofluorescence microscopy was performed on rat jejunum over a time course following intraluminal injection (ILI) of AMT-101. Co-localization of the human interleukin (IL)-10 element of AMT-101 with TMEM132A (a, c) or GRP75 (b, d) at 1- or 5-min post ILI. Apical (luminal) epithelial membrane (open arrow); basal epithelial cell surface-basement membrane demarcation (dashed line); lamina propria ( l-p ); goblet cell (g). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue).

Figure Legend Snippet: GRP75 associates with cholix in apical endosomes. Immunofluorescence microscopy was performed on rat jejunum over a time course following intraluminal injection (ILI) of AMT-101. Co-localization of the human interleukin (IL)-10 element of AMT-101 with TMEM132A (a, c) or GRP75 (b, d) at 1- or 5-min post ILI. Apical (luminal) epithelial membrane (open arrow); basal epithelial cell surface-basement membrane demarcation (dashed line); lamina propria ( l-p ); goblet cell (g). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue).

Techniques Used: Immunofluorescence, Microscopy, Injection, Membrane, Staining

Relationship of GRP75 to GRP78 and TMEM132A. Immunofluorescence microscopy was performed on rat jejunum over a time course following intraluminal injection (ILI) of Chx266-hGH. Co-localization events detecting the human growth hormone (hGH) element of Chx266-hGH with a, TMEM132A or b, GRP78 compared to the co-localization of c, TMEM132A and GRP78 at 5 min post ILI of Chx266-hGH. Apical (luminal) epithelial membrane (open arrow); basal epithelial cell surface-basement membrane demarcation (dashed line); lamina propria ( l-p ); goblet cell (g). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue).

Figure Legend Snippet: Relationship of GRP75 to GRP78 and TMEM132A. Immunofluorescence microscopy was performed on rat jejunum over a time course following intraluminal injection (ILI) of Chx266-hGH. Co-localization events detecting the human growth hormone (hGH) element of Chx266-hGH with a, TMEM132A or b, GRP78 compared to the co-localization of c, TMEM132A and GRP78 at 5 min post ILI of Chx266-hGH. Apical (luminal) epithelial membrane (open arrow); basal epithelial cell surface-basement membrane demarcation (dashed line); lamina propria ( l-p ); goblet cell (g). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue).

Techniques Used: Immunofluorescence, Microscopy, Injection, Membrane, Staining

GRP75 does not associate with cholix in basal vesicular structures. Immunofluorescence microscopy was performed on rat jejunum after intraluminal injection (ILI) of Chx266-hGH. The human growth hormone (hGH) element of Chx266-hGH was co-localized with early endosomal antigen 1 (EEA1; a, c) or GRP75 (b, d) at 5- or 15-min post ILI. Apical (luminal) epithelial membrane (open arrow); basal epithelial cell surface-basement membrane demarcation (dashed line); lamina propria ( l-p ); goblet cell (g). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue).

Figure Legend Snippet: GRP75 does not associate with cholix in basal vesicular structures. Immunofluorescence microscopy was performed on rat jejunum after intraluminal injection (ILI) of Chx266-hGH. The human growth hormone (hGH) element of Chx266-hGH was co-localized with early endosomal antigen 1 (EEA1; a, c) or GRP75 (b, d) at 5- or 15-min post ILI. Apical (luminal) epithelial membrane (open arrow); basal epithelial cell surface-basement membrane demarcation (dashed line); lamina propria ( l-p ); goblet cell (g). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue).

Techniques Used: Immunofluorescence, Microscopy, Injection, Membrane, Staining

Cholix trafficking involves LMAN1-positive compartments. Immunofluorescence microscopy was performed on rat jejunum after intraluminal injection (ILI) of Chx266-hGH. The human growth hormone (hGH) element of Chx266-hGH was co-localized with a, early endosomal antigen 1 (EEA1) or b, LMAN1 at 5 min post ILI. c, Co-localization of EEA1 and LMAN1 at 5 min post ILI. Co-localization of hGH with d, GRP75 or e, LMAN1 at 15 min post ILI of Chx266-hGH. f, Co-localization of GRP75 and LMAN1 at 15 min post ILI of Chx266-hGH. Apical (luminal) epithelial membrane (open arrow); basal epithelial cell surface-basement membrane demarcation (dashed line); lamina propria ( l-p ); goblet cell (g). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue).

Figure Legend Snippet: Cholix trafficking involves LMAN1-positive compartments. Immunofluorescence microscopy was performed on rat jejunum after intraluminal injection (ILI) of Chx266-hGH. The human growth hormone (hGH) element of Chx266-hGH was co-localized with a, early endosomal antigen 1 (EEA1) or b, LMAN1 at 5 min post ILI. c, Co-localization of EEA1 and LMAN1 at 5 min post ILI. Co-localization of hGH with d, GRP75 or e, LMAN1 at 15 min post ILI of Chx266-hGH. f, Co-localization of GRP75 and LMAN1 at 15 min post ILI of Chx266-hGH. Apical (luminal) epithelial membrane (open arrow); basal epithelial cell surface-basement membrane demarcation (dashed line); lamina propria ( l-p ); goblet cell (g). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue).

Techniques Used: Immunofluorescence, Microscopy, Injection, Membrane, Staining

GRP75 intersects with furin differently than with LMAN1. Immunofluorescence microscopy was performed on rat jejunum after intraluminal injection (ILI) of Chx266-hGH. The human growth hormone (hGH) element of Chx266-hGH was co-localized with a, furin or b, GRP75 at 15 min post ILI. c, Co-localization of furin and GRP75 at 15 min post ILI. Co-localization of hGH with d, furin or e, LMAN1 at 15 min post ILI of Chx266-hGH. f, Co-localization of furin and LMAN1 at 15 min post ILI of Chx266-hGH. Apical (luminal) epithelial membrane (open arrow); basal epithelial cell surface-basement membrane demarcation (dashed line); lamina propria ( l-p ); goblet cell (g). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue).

Figure Legend Snippet: GRP75 intersects with furin differently than with LMAN1. Immunofluorescence microscopy was performed on rat jejunum after intraluminal injection (ILI) of Chx266-hGH. The human growth hormone (hGH) element of Chx266-hGH was co-localized with a, furin or b, GRP75 at 15 min post ILI. c, Co-localization of furin and GRP75 at 15 min post ILI. Co-localization of hGH with d, furin or e, LMAN1 at 15 min post ILI of Chx266-hGH. f, Co-localization of furin and LMAN1 at 15 min post ILI of Chx266-hGH. Apical (luminal) epithelial membrane (open arrow); basal epithelial cell surface-basement membrane demarcation (dashed line); lamina propria ( l-p ); goblet cell (g). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue).

Techniques Used: Immunofluorescence, Microscopy, Injection, Membrane, Staining

Diagram of events associated with cholix apical to basal transcytosis with interaction partners. Highlighted steps are explained in the text with greater detail. (1) Cholix enters at the apical plasma membrane in the microvilli area where it interacts with TMEM132A and furin. (2) Cholix enters an early endosomal compartment where it traffics preferentially to a late endosome rather than a recycling pathway. (3) Instead of continuing along the lysosomal degradation pathway, cholix interacts with GRP75 and moves to a sorting endosome. (4) Reorganization of LMAN1 from the endoplasmic reticulum intermediate Golgi complex (ERGIC) to apical sorting vesicles. (5) In this location, LMAN1 can intersect with cholix delivered to this site with furin and GRP75. (6) LMAN1 and furin appear to return with cholix to a supranuclear region of the cell consistent with location of the ERGIC. (7) Reorganization of LMAN1 from the ERGIC to the basal vesicular compartment provides a route for cholix to this region of the cell. (8) Cholix in complex with LMAN1 enters sorting endosomes in the basal vesicular compartment. (9) Within basal sorting endosomes, cholix/LMAN1 intersects with perlecan in a recycling endosome, which delivers it to the basal plasma membrane, resulting in cholix exocytosis and completion of the apical to basal transcytosis process. (10) Perlecan recycles to the basal sorting endosome where it can engage cholix trafficked to this site with LMAN1. (11) Unlike in the apical vesicular compartment, GRP75 and furin present in the basal vesicular compartment are not associated with cholix in this region of enterocytes.

Figure Legend Snippet: Diagram of events associated with cholix apical to basal transcytosis with interaction partners. Highlighted steps are explained in the text with greater detail. (1) Cholix enters at the apical plasma membrane in the microvilli area where it interacts with TMEM132A and furin. (2) Cholix enters an early endosomal compartment where it traffics preferentially to a late endosome rather than a recycling pathway. (3) Instead of continuing along the lysosomal degradation pathway, cholix interacts with GRP75 and moves to a sorting endosome. (4) Reorganization of LMAN1 from the endoplasmic reticulum intermediate Golgi complex (ERGIC) to apical sorting vesicles. (5) In this location, LMAN1 can intersect with cholix delivered to this site with furin and GRP75. (6) LMAN1 and furin appear to return with cholix to a supranuclear region of the cell consistent with location of the ERGIC. (7) Reorganization of LMAN1 from the ERGIC to the basal vesicular compartment provides a route for cholix to this region of the cell. (8) Cholix in complex with LMAN1 enters sorting endosomes in the basal vesicular compartment. (9) Within basal sorting endosomes, cholix/LMAN1 intersects with perlecan in a recycling endosome, which delivers it to the basal plasma membrane, resulting in cholix exocytosis and completion of the apical to basal transcytosis process. (10) Perlecan recycles to the basal sorting endosome where it can engage cholix trafficked to this site with LMAN1. (11) Unlike in the apical vesicular compartment, GRP75 and furin present in the basal vesicular compartment are not associated with cholix in this region of enterocytes.

Techniques Used: Clinical Proteomics, Membrane

Western Blot:

Article Title: GRP75 as a functional element of cholix transcytosis
Article Snippet: .. Western blotting was performed using rabbit anti-human GRP75 as primary antibody followed by using Cy3-labeled Donkey anti-rabbit secondary antibody (Jackson ImmunoResearch, 1;2500). .. Western blot images were scanned using Typhoon 9400 (Applied Biomics, Hayward, CA).

Image Search Results

Journal: Tissue Barriers

Article Title: GRP75 as a functional element of cholix transcytosis

doi: 10.1080/21688370.2022.2039003

Figure Lengend Snippet: GRP75 is an interactive partner of cholix. Polarized monolayers of Caco-2 cells were exposed apically to nontoxic full-length Chx (ntChx) anchored through a C-terminal hexa-histidine sequence to 100 nm diameter magnetic beads. After 15 min, cell membranes were isolated and separated into non-magnetic and magnetic fractions. a, 1-D SDS-PAGE gel of non-magnetic and magnetic captured membrane fractions. b, 2-D gel separation of proteins present in non-magnetic and magnetic capture membrane fractions with location of spot 16 identified by mass spectrometry as GRP75. c, Pull-down (PD) using ntChx probed with antibodies to either GRP75 or human growth hormone (hGH). d, Immunoprecipitation of Caco-2 cell lysate using ntChx probed with anti-GRP75 or control antibodies. e, 2-D Western blot of magnetic capture membrane fraction probed with anti-GRP75. f, ELISA-based, neutral pH, format where bound ntChx or the control protein bovine serum albumin (BSA) was used to capture GRP75, heat shock protein 60 (HSP60), glucose-regulated protein 78 (GRP78), or HSP90.

Article Snippet: Western blotting was performed using rabbit anti-human GRP75 as primary antibody followed by using Cy3-labeled Donkey anti-rabbit secondary antibody (Jackson ImmunoResearch, 1;2500).

Techniques: Sequencing, Magnetic Beads, Isolation, SDS Page, Membrane, Mass Spectrometry, Immunoprecipitation, Control, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Tissue Barriers

Article Title: GRP75 as a functional element of cholix transcytosis

doi: 10.1080/21688370.2022.2039003

Figure Lengend Snippet: GRP75 and perlecan are functionally involved in cholix transcytosis. Expression of three proteins identified by ntChx-magnetic bead capture were suppressed using a CRISPR/cas 9-mediated knockdown protocol that targeted either GRP75, HSPG2 , or KRT8 in Caco-2 cells. Confluent monolayers of the resulting cell lines having decreased levels of a, GRP75, b, perlecan, or c, cytokeratin 8 (CT8) were evaluated for their capacity for Chx-mediated transcytosis using Chx266-hGH and for the nonspecific movement of the control protein hGH over a 60 min time period. d, Transcytosis of Chx266-hGH across confluent monolayers of parent Caco-2 cells was evaluated following a simultaneous apical application of a monoclonal antibody against GRP75 or perlecan, with an isotype antibody recognizing interleukin (IL)-10 serving as a control. Densitometry measurements of individual Western blot bands are shown for graphical comparison.

Techniques: Expressing, CRISPR, Knockdown, Control, Western Blot, Comparison

Journal: Tissue Barriers

Article Title: GRP75 as a functional element of cholix transcytosis

doi: 10.1080/21688370.2022.2039003

Figure Lengend Snippet: GRP75 associates with cholix in apical endosomes. Immunofluorescence microscopy was performed on rat jejunum over a time course following intraluminal injection (ILI) of AMT-101. Co-localization of the human interleukin (IL)-10 element of AMT-101 with TMEM132A (a, c) or GRP75 (b, d) at 1- or 5-min post ILI. Apical (luminal) epithelial membrane (open arrow); basal epithelial cell surface-basement membrane demarcation (dashed line); lamina propria ( l-p ); goblet cell (g). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue).

Techniques: Immunofluorescence, Microscopy, Injection, Membrane, Staining

Journal: Tissue Barriers

Article Title: GRP75 as a functional element of cholix transcytosis

doi: 10.1080/21688370.2022.2039003

Figure Lengend Snippet: Relationship of GRP75 to GRP78 and TMEM132A. Immunofluorescence microscopy was performed on rat jejunum over a time course following intraluminal injection (ILI) of Chx266-hGH. Co-localization events detecting the human growth hormone (hGH) element of Chx266-hGH with a, TMEM132A or b, GRP78 compared to the co-localization of c, TMEM132A and GRP78 at 5 min post ILI of Chx266-hGH. Apical (luminal) epithelial membrane (open arrow); basal epithelial cell surface-basement membrane demarcation (dashed line); lamina propria ( l-p ); goblet cell (g). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue).

Techniques: Immunofluorescence, Microscopy, Injection, Membrane, Staining

Journal: Tissue Barriers

Article Title: GRP75 as a functional element of cholix transcytosis

doi: 10.1080/21688370.2022.2039003

Figure Lengend Snippet: GRP75 does not associate with cholix in basal vesicular structures. Immunofluorescence microscopy was performed on rat jejunum after intraluminal injection (ILI) of Chx266-hGH. The human growth hormone (hGH) element of Chx266-hGH was co-localized with early endosomal antigen 1 (EEA1; a, c) or GRP75 (b, d) at 5- or 15-min post ILI. Apical (luminal) epithelial membrane (open arrow); basal epithelial cell surface-basement membrane demarcation (dashed line); lamina propria ( l-p ); goblet cell (g). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue).

Techniques: Immunofluorescence, Microscopy, Injection, Membrane, Staining

Journal: Tissue Barriers

Article Title: GRP75 as a functional element of cholix transcytosis

doi: 10.1080/21688370.2022.2039003

Figure Lengend Snippet: Cholix trafficking involves LMAN1-positive compartments. Immunofluorescence microscopy was performed on rat jejunum after intraluminal injection (ILI) of Chx266-hGH. The human growth hormone (hGH) element of Chx266-hGH was co-localized with a, early endosomal antigen 1 (EEA1) or b, LMAN1 at 5 min post ILI. c, Co-localization of EEA1 and LMAN1 at 5 min post ILI. Co-localization of hGH with d, GRP75 or e, LMAN1 at 15 min post ILI of Chx266-hGH. f, Co-localization of GRP75 and LMAN1 at 15 min post ILI of Chx266-hGH. Apical (luminal) epithelial membrane (open arrow); basal epithelial cell surface-basement membrane demarcation (dashed line); lamina propria ( l-p ); goblet cell (g). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue).

Techniques: Immunofluorescence, Microscopy, Injection, Membrane, Staining

Journal: Tissue Barriers

Article Title: GRP75 as a functional element of cholix transcytosis

doi: 10.1080/21688370.2022.2039003

Figure Lengend Snippet: GRP75 intersects with furin differently than with LMAN1. Immunofluorescence microscopy was performed on rat jejunum after intraluminal injection (ILI) of Chx266-hGH. The human growth hormone (hGH) element of Chx266-hGH was co-localized with a, furin or b, GRP75 at 15 min post ILI. c, Co-localization of furin and GRP75 at 15 min post ILI. Co-localization of hGH with d, furin or e, LMAN1 at 15 min post ILI of Chx266-hGH. f, Co-localization of furin and LMAN1 at 15 min post ILI of Chx266-hGH. Apical (luminal) epithelial membrane (open arrow); basal epithelial cell surface-basement membrane demarcation (dashed line); lamina propria ( l-p ); goblet cell (g). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue).

Techniques: Immunofluorescence, Microscopy, Injection, Membrane, Staining

Journal: Tissue Barriers

Article Title: GRP75 as a functional element of cholix transcytosis

doi: 10.1080/21688370.2022.2039003

Figure Lengend Snippet: Diagram of events associated with cholix apical to basal transcytosis with interaction partners. Highlighted steps are explained in the text with greater detail. (1) Cholix enters at the apical plasma membrane in the microvilli area where it interacts with TMEM132A and furin. (2) Cholix enters an early endosomal compartment where it traffics preferentially to a late endosome rather than a recycling pathway. (3) Instead of continuing along the lysosomal degradation pathway, cholix interacts with GRP75 and moves to a sorting endosome. (4) Reorganization of LMAN1 from the endoplasmic reticulum intermediate Golgi complex (ERGIC) to apical sorting vesicles. (5) In this location, LMAN1 can intersect with cholix delivered to this site with furin and GRP75. (6) LMAN1 and furin appear to return with cholix to a supranuclear region of the cell consistent with location of the ERGIC. (7) Reorganization of LMAN1 from the ERGIC to the basal vesicular compartment provides a route for cholix to this region of the cell. (8) Cholix in complex with LMAN1 enters sorting endosomes in the basal vesicular compartment. (9) Within basal sorting endosomes, cholix/LMAN1 intersects with perlecan in a recycling endosome, which delivers it to the basal plasma membrane, resulting in cholix exocytosis and completion of the apical to basal transcytosis process. (10) Perlecan recycles to the basal sorting endosome where it can engage cholix trafficked to this site with LMAN1. (11) Unlike in the apical vesicular compartment, GRP75 and furin present in the basal vesicular compartment are not associated with cholix in this region of enterocytes.

Techniques: Clinical Proteomics, Membrane

Association of mortalin, Bax or Bcl-2 expression with clinicopathological characteristics in 116 patients with intrahepatic cholangiocarcinoma.

Journal: Oncology Letters

Article Title: Characterization and prognostic significance of mortalin, Bcl-2 and Bax in intrahepatic cholangiocarcinoma

doi: 10.3892/ol.2017.7570

Figure Lengend Snippet: Association of mortalin, Bax or Bcl-2 expression with clinicopathological characteristics in 116 patients with intrahepatic cholangiocarcinoma.

Article Snippet: Rabbit anti-human mortalin monoclonal antibody (cat. no. 3593, dilution 1:100; Cell Signaling Technology, Inc., Danvers, MA, USA), mouse anti-human Bcl-2 monoclonal antibody (cat. no. ab182858, dilution 1:400) and mouse anti-human Bax monoclonal antibody (cat. no. ab77566; dilution 1:400) (both from Abcam, Cambridge, UK) were used to detect the expression of mortalin, Bcl-2 and Bax in ICC tissues, respectively.

Techniques: Expressing, Staining, Encapsulation

Expression of mortalin, Bcl-2 and Bax in ICC tissues and peritumoral samples. Representative images for H&E staining, and (A) mortalin, (B) Bcl-2 and (C) Bax immunohistochemical staining in tumor and peritumoral samples (scale bars, 200 µm). (D) A histogram demonstrating that the expression of mortalin in ICC tissues was high compared with peritumoral tissues (P<0.001). (E) The expression of Bcl-2 in ICC tissues was high compared with peritumoral tissues (P<0.001); and (F) that the expression of Bax in tumor samples was significantly lower than that in the peritumoral samples (P<0.001). Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-like protein 4; ICC, intrahepatic cholangiocarcinoma; H&E, hematoxylin and eosin.

Journal: Oncology Letters

Article Title: Characterization and prognostic significance of mortalin, Bcl-2 and Bax in intrahepatic cholangiocarcinoma

doi: 10.3892/ol.2017.7570

Figure Lengend Snippet: Expression of mortalin, Bcl-2 and Bax in ICC tissues and peritumoral samples. Representative images for H&E staining, and (A) mortalin, (B) Bcl-2 and (C) Bax immunohistochemical staining in tumor and peritumoral samples (scale bars, 200 µm). (D) A histogram demonstrating that the expression of mortalin in ICC tissues was high compared with peritumoral tissues (P<0.001). (E) The expression of Bcl-2 in ICC tissues was high compared with peritumoral tissues (P<0.001); and (F) that the expression of Bax in tumor samples was significantly lower than that in the peritumoral samples (P<0.001). Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-like protein 4; ICC, intrahepatic cholangiocarcinoma; H&E, hematoxylin and eosin.

Techniques: Expressing, Staining, Immunohistochemical staining

Associations between mortalin, Bcl-2 and Bax expression in ICC tumor samples. (A) Representative staining images demonstrating high moratlin and Bcl-2 expression and low Bax expression tissue (case 3), and low mortalin and Bcl-2 expression and low Bax expression tissue (case 9) (scale bars, 200 µm). Positive correlations were detected between (B) mortalin and Bcl-2 expression in ICC tissues (r=0.398, P<0.001); (C) between mortalin and Bax expression (r=−0.238, P=0.010) and (D) between Bcl-2 and Bax expression (r=−0.207, P=0.026). Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-like protein 4; ICC, intrahepatic cholangiocarcinoma; H&E, hematoxylin and eosin.

Journal: Oncology Letters

Article Title: Characterization and prognostic significance of mortalin, Bcl-2 and Bax in intrahepatic cholangiocarcinoma

doi: 10.3892/ol.2017.7570

Figure Lengend Snippet: Associations between mortalin, Bcl-2 and Bax expression in ICC tumor samples. (A) Representative staining images demonstrating high moratlin and Bcl-2 expression and low Bax expression tissue (case 3), and low mortalin and Bcl-2 expression and low Bax expression tissue (case 9) (scale bars, 200 µm). Positive correlations were detected between (B) mortalin and Bcl-2 expression in ICC tissues (r=0.398, P<0.001); (C) between mortalin and Bax expression (r=−0.238, P=0.010) and (D) between Bcl-2 and Bax expression (r=−0.207, P=0.026). Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-like protein 4; ICC, intrahepatic cholangiocarcinoma; H&E, hematoxylin and eosin.

Techniques: Expressing, Staining

Univariate and multivariate Cox regression analysis of factors associated with overall survival rate and cumulative recurrence rate.

Journal: Oncology Letters

Article Title: Characterization and prognostic significance of mortalin, Bcl-2 and Bax in intrahepatic cholangiocarcinoma

doi: 10.3892/ol.2017.7570

Figure Lengend Snippet: Univariate and multivariate Cox regression analysis of factors associated with overall survival rate and cumulative recurrence rate.

Techniques: Encapsulation, Staining

Kaplan-Meier analysis and log-rank test analysis of prognostic significance. Representative images of H&E and immunohistochemical staining for (A) mortalin, (B) Bcl-2 and (C) Bax in ICC samples (scale bars, 200 µm). ICC patients with high mortalin expression had a poorer prognosis according to (D) overall survival and (E) cumulative recurrence compared with those exhibiting low mortalin expression. ICC patients exhibiting high Bcl-2 expression had a poorer prognosis according to (F) overall survival and (G) cumulative recurrence compared with those exhibiting low Bcl-2 expression. ICC patients with low Bax expression had a poorer prognosis according to (H) overall survival and (I) cumulative recurrence compared with those exhibiting high Bax expression. Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-like protein 4; H&E, hematoxylin and eosin; ICC, intrahepatic cholangiocarcinoma.

Journal: Oncology Letters

Article Title: Characterization and prognostic significance of mortalin, Bcl-2 and Bax in intrahepatic cholangiocarcinoma

doi: 10.3892/ol.2017.7570

Figure Lengend Snippet: Kaplan-Meier analysis and log-rank test analysis of prognostic significance. Representative images of H&E and immunohistochemical staining for (A) mortalin, (B) Bcl-2 and (C) Bax in ICC samples (scale bars, 200 µm). ICC patients with high mortalin expression had a poorer prognosis according to (D) overall survival and (E) cumulative recurrence compared with those exhibiting low mortalin expression. ICC patients exhibiting high Bcl-2 expression had a poorer prognosis according to (F) overall survival and (G) cumulative recurrence compared with those exhibiting low Bcl-2 expression. ICC patients with low Bax expression had a poorer prognosis according to (H) overall survival and (I) cumulative recurrence compared with those exhibiting high Bax expression. Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-like protein 4; H&E, hematoxylin and eosin; ICC, intrahepatic cholangiocarcinoma.

Techniques: Immunohistochemical staining, Staining, Expressing

Combined expression of mortalin, Bcl-2 and Bax to predict ICC prognosis via Kaplan-Meier analysis. (A) Patients with ICC exhibiting high mortalin and Bcl-2 expression and low Bax expression had a relatively poor prognosis according to overall survival rates of the 3 subgroups: I, high mortalin and Bcl-2 expression and low Bax expression; II, low mortalin and Bcl-2 expression and high Bax expression; and III, all possible remaining combinations of expression of the 3 proteins. (B) Patients with ICC exhibiting high mortalin and Bcl-2 expression and low Bax expression were associated with the worst prognosis according to cumulative recurrence rates in the 3 subgroups. Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-like protein 4; ICC, intrahepatic cholangiocarcinoma.

Journal: Oncology Letters

Article Title: Characterization and prognostic significance of mortalin, Bcl-2 and Bax in intrahepatic cholangiocarcinoma

doi: 10.3892/ol.2017.7570

Figure Lengend Snippet: Combined expression of mortalin, Bcl-2 and Bax to predict ICC prognosis via Kaplan-Meier analysis. (A) Patients with ICC exhibiting high mortalin and Bcl-2 expression and low Bax expression had a relatively poor prognosis according to overall survival rates of the 3 subgroups: I, high mortalin and Bcl-2 expression and low Bax expression; II, low mortalin and Bcl-2 expression and high Bax expression; and III, all possible remaining combinations of expression of the 3 proteins. (B) Patients with ICC exhibiting high mortalin and Bcl-2 expression and low Bax expression were associated with the worst prognosis according to cumulative recurrence rates in the 3 subgroups. Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-like protein 4; ICC, intrahepatic cholangiocarcinoma.

Techniques: Expressing

Tumor tissue arrays containing 63 pairs of non-tumor and HCC tissues were stained with GRP75 and HSP90 specific antibodies using a DAB detection kit. (A, C) Representative images of immunohistochemically stained GRP75 or HSP90 proteins in paraffin-embedded non-tumor liver and liver tumor tissues. Normal and tumor tissues were classified into four groups based on staining intensities. (B, D) Tabulation of the percentage of normal, T2 and T3 cells within each group. 32 from pathologic stage T2 patients and 31 from T3 patients, tumor staging was determined according to the sixth edition of the TNM (tumor-node-metastasis, TNM) classification of International Union Against Cancer.

Journal: PLoS ONE

Article Title: Targeting GRP75 Improves HSP90 Inhibitor Efficacy by Enhancing p53-Mediated Apoptosis in Hepatocellular Carcinoma

doi: 10.1371/journal.pone.0085766

Figure Lengend Snippet: Tumor tissue arrays containing 63 pairs of non-tumor and HCC tissues were stained with GRP75 and HSP90 specific antibodies using a DAB detection kit. (A, C) Representative images of immunohistochemically stained GRP75 or HSP90 proteins in paraffin-embedded non-tumor liver and liver tumor tissues. Normal and tumor tissues were classified into four groups based on staining intensities. (B, D) Tabulation of the percentage of normal, T2 and T3 cells within each group. 32 from pathologic stage T2 patients and 31 from T3 patients, tumor staging was determined according to the sixth edition of the TNM (tumor-node-metastasis, TNM) classification of International Union Against Cancer.

Article Snippet: Tissue slides were incubated with rabbit anti-human GRP75 (1∶50; Cell Signaling Technology, Boston, MA, U.S.

Techniques: Staining

(A) The indicated cell lines were treated with MKT-077 (MKT) or 17-AAG alone or in combination at indicated dosages for 24 hours. Cell viability was qualified using the Cell Counting Kit-8 (Dojindo Laboratories). * denotes P < 0.05 comparing 17-AAG (1 µ M or 5 µ M) to MKT-077 (3 µ g/mL) +17-AAG (1 µ M or 5 µ M). (B) Increased cell apoptosis following combined treatment. Bel-7402, HuH7 and Hep3B cells treated as described above were subjected to Annexin-V and PI staining and cell apoptosis was quantified by FACS. * denotes P < 0.05 comparing 17-AAG (1 µ M or 5 µ M) to MKT-077 (3 µ g/mL) +17-AAG (1 µ M or 5 µ M). (C) Levels of cleaved PARP, GRP75, Akt, phospho-Akt Ser473 , phosphor-Akt Thr308 , p53, phospho-p53 Ser15 , and phospho-p53 Ser37 were detected by Western Blotting analysis. GAPDH served as a loading control.

Journal: PLoS ONE

Article Title: Targeting GRP75 Improves HSP90 Inhibitor Efficacy by Enhancing p53-Mediated Apoptosis in Hepatocellular Carcinoma

doi: 10.1371/journal.pone.0085766

Figure Lengend Snippet: (A) The indicated cell lines were treated with MKT-077 (MKT) or 17-AAG alone or in combination at indicated dosages for 24 hours. Cell viability was qualified using the Cell Counting Kit-8 (Dojindo Laboratories). * denotes P < 0.05 comparing 17-AAG (1 µ M or 5 µ M) to MKT-077 (3 µ g/mL) +17-AAG (1 µ M or 5 µ M). (B) Increased cell apoptosis following combined treatment. Bel-7402, HuH7 and Hep3B cells treated as described above were subjected to Annexin-V and PI staining and cell apoptosis was quantified by FACS. * denotes P < 0.05 comparing 17-AAG (1 µ M or 5 µ M) to MKT-077 (3 µ g/mL) +17-AAG (1 µ M or 5 µ M). (C) Levels of cleaved PARP, GRP75, Akt, phospho-Akt Ser473 , phosphor-Akt Thr308 , p53, phospho-p53 Ser15 , and phospho-p53 Ser37 were detected by Western Blotting analysis. GAPDH served as a loading control.

Article Snippet: Tissue slides were incubated with rabbit anti-human GRP75 (1∶50; Cell Signaling Technology, Boston, MA, U.S.

Techniques: Cell Counting, Staining, Western Blot, Control

$(A, B) Bel-7402 and HuH7 cells were treated with MKT-077, 17-AAG or a combination of both agents at the indicated concentrations for 24 hours. Cytoplasmic and nuclear fractions of Bel-7402 and HuH7 cells were separated, and subjected to Western blotting analysis of p53 expression. α-tublin and Lamin B1 served as loading controls for cytoplasmic and nuclear fractions respectively. (C) Hep3B cells infected with Lenti-p53 (Hep3B-p53 +/+ ) were treated with MKT-077, 17-AAG or MKT-077+17-AAG at indicated dosages for 24 hours, and immunofluorescent staining was performed to determine p53 subcellular localization. (D) Bel-7402 cells treated with MKT-077 or 17-AAG alone, or MKT-077+17-AAG for 24 hours were harvested and subjected to co-immunoprecipitation using GRP75-specific antibodies. Levels of p53, Akt and HSP90 were detected by Western blotting analysis. (E) MKT-077 and 17-AAG synergistically enhanced mRNA levels of the p53 target genes p21 and PUMA , and reduced MDM2 mRNA levels. The mRNA levels of these genes were quantified using real-time RT-PCR and normalized to levels of GAPDH mRNA. * denotes P < 0.05 comparing 17-AAG to MKT-077+17-AAG.$

Journal: PLoS ONE

Article Title: Targeting GRP75 Improves HSP90 Inhibitor Efficacy by Enhancing p53-Mediated Apoptosis in Hepatocellular Carcinoma

doi: 10.1371/journal.pone.0085766

Figure Lengend Snippet: (A, B) Bel-7402 and HuH7 cells were treated with MKT-077, 17-AAG or a combination of both agents at the indicated concentrations for 24 hours. Cytoplasmic and nuclear fractions of Bel-7402 and HuH7 cells were separated, and subjected to Western blotting analysis of p53 expression. α-tublin and Lamin B1 served as loading controls for cytoplasmic and nuclear fractions respectively. (C) Hep3B cells infected with Lenti-p53 (Hep3B-p53 +/+ ) were treated with MKT-077, 17-AAG or MKT-077+17-AAG at indicated dosages for 24 hours, and immunofluorescent staining was performed to determine p53 subcellular localization. (D) Bel-7402 cells treated with MKT-077 or 17-AAG alone, or MKT-077+17-AAG for 24 hours were harvested and subjected to co-immunoprecipitation using GRP75-specific antibodies. Levels of p53, Akt and HSP90 were detected by Western blotting analysis. (E) MKT-077 and 17-AAG synergistically enhanced mRNA levels of the p53 target genes p21 and PUMA , and reduced MDM2 mRNA levels. The mRNA levels of these genes were quantified using real-time RT-PCR and normalized to levels of GAPDH mRNA. * denotes P < 0.05 comparing 17-AAG to MKT-077+17-AAG.

Article Snippet: Tissue slides were incubated with rabbit anti-human GRP75 (1∶50; Cell Signaling Technology, Boston, MA, U.S.

Techniques: Western Blot, Expressing, Infection, Staining, Immunoprecipitation, Quantitative RT-PCR

Review

rabbit anti human grp75 (Jackson Immuno)

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1) Product Images from "GRP75 as a functional element of cholix transcytosis"

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