af244 Search Results


af244  (R&D Systems)
90
R&D Systems af244
ICAM-2 WT and variants co-precipitated with α-actinin. A ) IP/IB experiments with control cell lines demonstrate the expected association of ICAM-2, α-actinin and actin in lysates from SK-N-ASpIRES.ICAM-2 cells (labeled as WT), but not SK-N-ASpIRESneo cells (neo). Results for these control cell lines were published previously . B ) ICAM-2 glycosylation site variants associated simultaneously with α-actinin and actin. Immunoprecipitations (IP) were performed using whole cell lysates and a mouse monoclonal antibody to α-actinin (MAB1682, Millipore). Following protein separation by electrophoresis, immunoblots (IB) were performed with antibodies to α-actinin (sc-7454, Santa Cruz Biotech), ICAM-2 <t>(AF244,</t> R&D Systems), or actin (4968, Cell Signaling). The presence of ICAM-2 WT and variants in each preparation was confirmed by immunoblot analysis of input preparations (a representative blot is shown in Figure C) and also by immunoblot analysis of the proteins remaining in the supernatant following precipitation (not shown), to confirm the presence of sufficient/excess ICAM-2 protein or variant in each preparation used for immunoprecipitation experiments.
Af244, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/af244/product/R&D Systems
Average 90 stars, based on 1 article reviews
af244 - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier
α actinin  (R&D Systems)
86
R&D Systems α actinin
Transfected SK-N-AS cells express ICAM-2 transcripts and proteins. A ) RNA from control human dermal microvascular endothelial cells (HDMVEC) generated RT-PCR products of the predicted 631 base pairs. All ICAM-2 transfectants contained readily detectable ICAM-2 RNA. B ) Bar graph depicting quantitation of the RT-PCR products shown in “A”. RNA expression levels were within 20% among the transfectants. This level of variability was not statistically significant. C ) Immunoblots of whole cell lysates (40 μg protein/lane) demonstrated that control HBMEC-28 cells and WT transfectants expressed immunoreactive protein having an apparent molecular mass of 55-60 kDa. ICAM-2 variants expressed proteins of appropriately lesser apparent masses. Transfectants expressed equivalent levels of actin and <t>α-actinin.</t> D ) Deglycosylation of proteins in whole cell lysates and subsequent immunoblots for ICAM-2 confirmed that all variants displayed the expected apparent molecular mass of ~30kDa. The “nonspecific” band that appears at ~36kDa in all lanes marked + PNGase F is the PNGase F protein itself. E ) Quantitation of ICAM-2 variants (lanes indicated as + PNGase F) was done to compare relative amounts of ICAM-2 protein expressed by transfectants. Results were normalized to the level of actin expression for each cell line. F ) ICAM-2 WT and variants localized to cell membranes. Experimental details are included in Methods. G ) Membrane localization of ICAM-2 WT and gsv4,5 was confirmed by fluorescence activated cell sorting (FACS) of intact cells, incubated with an antibody recognizing the extracellular domain (ED) of ICAM-2 (CBR-IC2/2) and a PE-conjugated secondary antibody. Non-intact cells were gated out using light scatter parameters and propidium iodide uptake. FACS profiles shown are for PE-positive cells generated using negative control IgG (blue line) or anti-ICAM-2 (red line). H ) Biotinylation experiments also demonstrated that ICAM-2 WT and variants localized to cell membranes. Experimental details are included in Methods.
α Actinin, supplied by R&D Systems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/α actinin/product/R&D Systems
Average 86 stars, based on 1 article reviews
α actinin - by Bioz Stars, 2026-03
86/100 stars
  Buy from Supplier

Image Search Results


ICAM-2 WT and variants co-precipitated with α-actinin. A ) IP/IB experiments with control cell lines demonstrate the expected association of ICAM-2, α-actinin and actin in lysates from SK-N-ASpIRES.ICAM-2 cells (labeled as WT), but not SK-N-ASpIRESneo cells (neo). Results for these control cell lines were published previously . B ) ICAM-2 glycosylation site variants associated simultaneously with α-actinin and actin. Immunoprecipitations (IP) were performed using whole cell lysates and a mouse monoclonal antibody to α-actinin (MAB1682, Millipore). Following protein separation by electrophoresis, immunoblots (IB) were performed with antibodies to α-actinin (sc-7454, Santa Cruz Biotech), ICAM-2 (AF244, R&D Systems), or actin (4968, Cell Signaling). The presence of ICAM-2 WT and variants in each preparation was confirmed by immunoblot analysis of input preparations (a representative blot is shown in Figure C) and also by immunoblot analysis of the proteins remaining in the supernatant following precipitation (not shown), to confirm the presence of sufficient/excess ICAM-2 protein or variant in each preparation used for immunoprecipitation experiments.

Journal: BMC Cancer

Article Title: N-glycosylation of ICAM-2 is required for ICAM-2-mediated complete suppression of metastatic potential of SK-N-AS neuroblastoma cells

doi: 10.1186/1471-2407-13-261

Figure Lengend Snippet: ICAM-2 WT and variants co-precipitated with α-actinin. A ) IP/IB experiments with control cell lines demonstrate the expected association of ICAM-2, α-actinin and actin in lysates from SK-N-ASpIRES.ICAM-2 cells (labeled as WT), but not SK-N-ASpIRESneo cells (neo). Results for these control cell lines were published previously . B ) ICAM-2 glycosylation site variants associated simultaneously with α-actinin and actin. Immunoprecipitations (IP) were performed using whole cell lysates and a mouse monoclonal antibody to α-actinin (MAB1682, Millipore). Following protein separation by electrophoresis, immunoblots (IB) were performed with antibodies to α-actinin (sc-7454, Santa Cruz Biotech), ICAM-2 (AF244, R&D Systems), or actin (4968, Cell Signaling). The presence of ICAM-2 WT and variants in each preparation was confirmed by immunoblot analysis of input preparations (a representative blot is shown in Figure C) and also by immunoblot analysis of the proteins remaining in the supernatant following precipitation (not shown), to confirm the presence of sufficient/excess ICAM-2 protein or variant in each preparation used for immunoprecipitation experiments.

Article Snippet: Antibodies used for immunoblots (IB) for ICAM-2, α-actinin, and actin were AF244 (R&D Systems, Minneapolis, MN) and sc-1512 (Santa Cruz Biotech, Santa Cruz, CA), 4968 (Cell Signaling Technology, Danvers, MA), and A5316 (Sigma-Aldrich, St. Louis, MO), respectively.

Techniques: Control, Labeling, Glycoproteomics, Electrophoresis, Western Blot, Variant Assay, Immunoprecipitation

Transfected SK-N-AS cells express ICAM-2 transcripts and proteins. A ) RNA from control human dermal microvascular endothelial cells (HDMVEC) generated RT-PCR products of the predicted 631 base pairs. All ICAM-2 transfectants contained readily detectable ICAM-2 RNA. B ) Bar graph depicting quantitation of the RT-PCR products shown in “A”. RNA expression levels were within 20% among the transfectants. This level of variability was not statistically significant. C ) Immunoblots of whole cell lysates (40 μg protein/lane) demonstrated that control HBMEC-28 cells and WT transfectants expressed immunoreactive protein having an apparent molecular mass of 55-60 kDa. ICAM-2 variants expressed proteins of appropriately lesser apparent masses. Transfectants expressed equivalent levels of actin and α-actinin. D ) Deglycosylation of proteins in whole cell lysates and subsequent immunoblots for ICAM-2 confirmed that all variants displayed the expected apparent molecular mass of ~30kDa. The “nonspecific” band that appears at ~36kDa in all lanes marked + PNGase F is the PNGase F protein itself. E ) Quantitation of ICAM-2 variants (lanes indicated as + PNGase F) was done to compare relative amounts of ICAM-2 protein expressed by transfectants. Results were normalized to the level of actin expression for each cell line. F ) ICAM-2 WT and variants localized to cell membranes. Experimental details are included in Methods. G ) Membrane localization of ICAM-2 WT and gsv4,5 was confirmed by fluorescence activated cell sorting (FACS) of intact cells, incubated with an antibody recognizing the extracellular domain (ED) of ICAM-2 (CBR-IC2/2) and a PE-conjugated secondary antibody. Non-intact cells were gated out using light scatter parameters and propidium iodide uptake. FACS profiles shown are for PE-positive cells generated using negative control IgG (blue line) or anti-ICAM-2 (red line). H ) Biotinylation experiments also demonstrated that ICAM-2 WT and variants localized to cell membranes. Experimental details are included in Methods.

Journal: BMC Cancer

Article Title: N-glycosylation of ICAM-2 is required for ICAM-2-mediated complete suppression of metastatic potential of SK-N-AS neuroblastoma cells

doi: 10.1186/1471-2407-13-261

Figure Lengend Snippet: Transfected SK-N-AS cells express ICAM-2 transcripts and proteins. A ) RNA from control human dermal microvascular endothelial cells (HDMVEC) generated RT-PCR products of the predicted 631 base pairs. All ICAM-2 transfectants contained readily detectable ICAM-2 RNA. B ) Bar graph depicting quantitation of the RT-PCR products shown in “A”. RNA expression levels were within 20% among the transfectants. This level of variability was not statistically significant. C ) Immunoblots of whole cell lysates (40 μg protein/lane) demonstrated that control HBMEC-28 cells and WT transfectants expressed immunoreactive protein having an apparent molecular mass of 55-60 kDa. ICAM-2 variants expressed proteins of appropriately lesser apparent masses. Transfectants expressed equivalent levels of actin and α-actinin. D ) Deglycosylation of proteins in whole cell lysates and subsequent immunoblots for ICAM-2 confirmed that all variants displayed the expected apparent molecular mass of ~30kDa. The “nonspecific” band that appears at ~36kDa in all lanes marked + PNGase F is the PNGase F protein itself. E ) Quantitation of ICAM-2 variants (lanes indicated as + PNGase F) was done to compare relative amounts of ICAM-2 protein expressed by transfectants. Results were normalized to the level of actin expression for each cell line. F ) ICAM-2 WT and variants localized to cell membranes. Experimental details are included in Methods. G ) Membrane localization of ICAM-2 WT and gsv4,5 was confirmed by fluorescence activated cell sorting (FACS) of intact cells, incubated with an antibody recognizing the extracellular domain (ED) of ICAM-2 (CBR-IC2/2) and a PE-conjugated secondary antibody. Non-intact cells were gated out using light scatter parameters and propidium iodide uptake. FACS profiles shown are for PE-positive cells generated using negative control IgG (blue line) or anti-ICAM-2 (red line). H ) Biotinylation experiments also demonstrated that ICAM-2 WT and variants localized to cell membranes. Experimental details are included in Methods.

Article Snippet: Antibodies used for immunoblots (IB) for ICAM-2, α-actinin, and actin were AF244 (R&D Systems, Minneapolis, MN) and sc-1512 (Santa Cruz Biotech, Santa Cruz, CA), 4968 (Cell Signaling Technology, Danvers, MA), and A5316 (Sigma-Aldrich, St. Louis, MO), respectively.

Techniques: Transfection, Generated, Reverse Transcription Polymerase Chain Reaction, Quantitation Assay, RNA Expression, Western Blot, Expressing, Membrane, Fluorescence, FACS, Incubation, Negative Control

ICAM-2 WT and variants co-precipitated with α-actinin. A ) IP/IB experiments with control cell lines demonstrate the expected association of ICAM-2, α-actinin and actin in lysates from SK-N-ASpIRES.ICAM-2 cells (labeled as WT), but not SK-N-ASpIRESneo cells (neo). Results for these control cell lines were published previously . B ) ICAM-2 glycosylation site variants associated simultaneously with α-actinin and actin. Immunoprecipitations (IP) were performed using whole cell lysates and a mouse monoclonal antibody to α-actinin (MAB1682, Millipore). Following protein separation by electrophoresis, immunoblots (IB) were performed with antibodies to α-actinin (sc-7454, Santa Cruz Biotech), ICAM-2 (AF244, R&D Systems), or actin (4968, Cell Signaling). The presence of ICAM-2 WT and variants in each preparation was confirmed by immunoblot analysis of input preparations (a representative blot is shown in Figure C) and also by immunoblot analysis of the proteins remaining in the supernatant following precipitation (not shown), to confirm the presence of sufficient/excess ICAM-2 protein or variant in each preparation used for immunoprecipitation experiments.

Journal: BMC Cancer

Article Title: N-glycosylation of ICAM-2 is required for ICAM-2-mediated complete suppression of metastatic potential of SK-N-AS neuroblastoma cells

doi: 10.1186/1471-2407-13-261

Figure Lengend Snippet: ICAM-2 WT and variants co-precipitated with α-actinin. A ) IP/IB experiments with control cell lines demonstrate the expected association of ICAM-2, α-actinin and actin in lysates from SK-N-ASpIRES.ICAM-2 cells (labeled as WT), but not SK-N-ASpIRESneo cells (neo). Results for these control cell lines were published previously . B ) ICAM-2 glycosylation site variants associated simultaneously with α-actinin and actin. Immunoprecipitations (IP) were performed using whole cell lysates and a mouse monoclonal antibody to α-actinin (MAB1682, Millipore). Following protein separation by electrophoresis, immunoblots (IB) were performed with antibodies to α-actinin (sc-7454, Santa Cruz Biotech), ICAM-2 (AF244, R&D Systems), or actin (4968, Cell Signaling). The presence of ICAM-2 WT and variants in each preparation was confirmed by immunoblot analysis of input preparations (a representative blot is shown in Figure C) and also by immunoblot analysis of the proteins remaining in the supernatant following precipitation (not shown), to confirm the presence of sufficient/excess ICAM-2 protein or variant in each preparation used for immunoprecipitation experiments.

Article Snippet: Antibodies used for immunoblots (IB) for ICAM-2, α-actinin, and actin were AF244 (R&D Systems, Minneapolis, MN) and sc-1512 (Santa Cruz Biotech, Santa Cruz, CA), 4968 (Cell Signaling Technology, Danvers, MA), and A5316 (Sigma-Aldrich, St. Louis, MO), respectively.

Techniques: Labeling, Electrophoresis, Western Blot, Variant Assay, Immunoprecipitation