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criterion tgx stain  (Bio-Rad)


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    Structured Review

    Bio-Rad criterion tgx stain
    Relative HSP abundances in whole skeletal muscle homogenates from young adults and older adults pre and post HIT exercise. Representative Westen blots of (A) HSP72, HSP27, and αB-crystallin and (B) phosphorylated HSP27 Ser15 (pHSP27 Ser15) and pαB-crystallin Ser59 in whole muscle homogenates from the vastus lateralis of the same individuals. Calibration curves of mixed muscle homogenates are indicated and were used to determine the relative number of given proteins (see <t>Methods).</t> <t>Stain-free</t> gels are indicative of total protein loading, and molecular weights are indicated by markers collected under white light capture without moving the membrane between that and chemiluminescence detection. Relative abundances of (C) HSP72, (D) HSP27, (E) pHSP27 Ser15, (F) αB-crystallin, and (G) pαB-crystallin Ser59 from young (circle) and older adults Pre (square) and older adults Post (triangle) HIT exercise are shown relative to average Old (pre) on a given gel (data are presented as mean ± SD). Individuals indicated by the number of symbols ( n : 5–7), with the same color assigned to the same individual and consistent across all graphs. * p ≤ 0.05 indicates Brown-Forsye and Welch’s and post hoc analysis using Games-Horwell. HIT = high-intensity training; HSP = heat shock protein; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.
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    Images

    1) Product Images from "Exercise attenuates stress-related signaling as sensed by higher phosphorylation of small heat shock proteins in skeletal muscle from older individuals"

    Article Title: Exercise attenuates stress-related signaling as sensed by higher phosphorylation of small heat shock proteins in skeletal muscle from older individuals

    Journal: Journal of Sport and Health Science

    doi: 10.1016/j.jshs.2025.101111

    Relative HSP abundances in whole skeletal muscle homogenates from young adults and older adults pre and post HIT exercise. Representative Westen blots of (A) HSP72, HSP27, and αB-crystallin and (B) phosphorylated HSP27 Ser15 (pHSP27 Ser15) and pαB-crystallin Ser59 in whole muscle homogenates from the vastus lateralis of the same individuals. Calibration curves of mixed muscle homogenates are indicated and were used to determine the relative number of given proteins (see Methods). Stain-free gels are indicative of total protein loading, and molecular weights are indicated by markers collected under white light capture without moving the membrane between that and chemiluminescence detection. Relative abundances of (C) HSP72, (D) HSP27, (E) pHSP27 Ser15, (F) αB-crystallin, and (G) pαB-crystallin Ser59 from young (circle) and older adults Pre (square) and older adults Post (triangle) HIT exercise are shown relative to average Old (pre) on a given gel (data are presented as mean ± SD). Individuals indicated by the number of symbols ( n : 5–7), with the same color assigned to the same individual and consistent across all graphs. * p ≤ 0.05 indicates Brown-Forsye and Welch’s and post hoc analysis using Games-Horwell. HIT = high-intensity training; HSP = heat shock protein; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.
    Figure Legend Snippet: Relative HSP abundances in whole skeletal muscle homogenates from young adults and older adults pre and post HIT exercise. Representative Westen blots of (A) HSP72, HSP27, and αB-crystallin and (B) phosphorylated HSP27 Ser15 (pHSP27 Ser15) and pαB-crystallin Ser59 in whole muscle homogenates from the vastus lateralis of the same individuals. Calibration curves of mixed muscle homogenates are indicated and were used to determine the relative number of given proteins (see Methods). Stain-free gels are indicative of total protein loading, and molecular weights are indicated by markers collected under white light capture without moving the membrane between that and chemiluminescence detection. Relative abundances of (C) HSP72, (D) HSP27, (E) pHSP27 Ser15, (F) αB-crystallin, and (G) pαB-crystallin Ser59 from young (circle) and older adults Pre (square) and older adults Post (triangle) HIT exercise are shown relative to average Old (pre) on a given gel (data are presented as mean ± SD). Individuals indicated by the number of symbols ( n : 5–7), with the same color assigned to the same individual and consistent across all graphs. * p ≤ 0.05 indicates Brown-Forsye and Welch’s and post hoc analysis using Games-Horwell. HIT = high-intensity training; HSP = heat shock protein; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.

    Techniques Used: Staining, Membrane

    HSP abundances in type I and II skeletal muscle fibers from young and older adults. (A, C, and F) The MHC isoform present was determined in individual muscle fiber segments from the vastus lateralis and, following pooling into type I and type II groups from a given biopsy, were analyzed by Westen blotting. Westen blots of (A) HSP72, (C) HSP27 and pHSP27 Ser15, (F) αB-crystallin and pαB-crystallin Ser59, with MHC isoforms in groups of fibers. Stain-free gels are indicative of total protein loading, and molecular weights are indicated by markers collected under white light capture without moving the membrane between that and chemiluminescence detection. Calibration curves of mixed muscle homogenates are indicated. Relative protein abundances of (B) HSP72, (D) HSP27, (E) pHSP27 Ser15, (G) αB-crystallin, and (H) pαB-crystallin Ser59 in fibers from young (circle) and older adults (square) type I fibers (no outline) and type II fibers (outline). All fibers are expressed relative to the average older adult’s type I fibers. The same color is assigned to the same individual and is consistent with (data are presented as mean ± SD). * p < 0.05 and ** p < 0.01, mixed effect model Univariant using either Tukey’s or Games-Horwell’s multiple comparison test (see Methods). HIT = high-intensity training; HSP = heat shock protein; MHC = myosin heavy chain; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.
    Figure Legend Snippet: HSP abundances in type I and II skeletal muscle fibers from young and older adults. (A, C, and F) The MHC isoform present was determined in individual muscle fiber segments from the vastus lateralis and, following pooling into type I and type II groups from a given biopsy, were analyzed by Westen blotting. Westen blots of (A) HSP72, (C) HSP27 and pHSP27 Ser15, (F) αB-crystallin and pαB-crystallin Ser59, with MHC isoforms in groups of fibers. Stain-free gels are indicative of total protein loading, and molecular weights are indicated by markers collected under white light capture without moving the membrane between that and chemiluminescence detection. Calibration curves of mixed muscle homogenates are indicated. Relative protein abundances of (B) HSP72, (D) HSP27, (E) pHSP27 Ser15, (G) αB-crystallin, and (H) pαB-crystallin Ser59 in fibers from young (circle) and older adults (square) type I fibers (no outline) and type II fibers (outline). All fibers are expressed relative to the average older adult’s type I fibers. The same color is assigned to the same individual and is consistent with (data are presented as mean ± SD). * p < 0.05 and ** p < 0.01, mixed effect model Univariant using either Tukey’s or Games-Horwell’s multiple comparison test (see Methods). HIT = high-intensity training; HSP = heat shock protein; MHC = myosin heavy chain; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.

    Techniques Used: Staining, Membrane, Comparison

    Related Articles

    Membrane:

    Article Title: FMRP-dependent translational control negatively regulates adapter protein complex 2-mediated endocytosis
    Article Snippet: Gel was hydrated by MilliQ water for 30min before imaged by ChemDoc Imaging Systems (Bio-Rad). .. Samples were run on a 4–20% Criterion TGX gel (Bio-Rad, 4561093) and transferred onto 0.2μm Nitrocellulose membrane (Bio-Rad, 1620115). ..

    Article Title: STING causes replication stress and nascent DNA degradation via SAMHD1
    Article Snippet: .. Equal protein amounts were separated by SDS-PAGE on a 4-15% Criterion TGX Gel (Bio-Rad) followed by transferring to a nitrocellulose membrane using the Trans-Blot Turbo system (Bio-Rad). ..

    Bradford Assay:

    Article Title:
    Article Snippet: .. Same amounts of lysate, adjusted based on the total protein levels measured by Bradford assay, were subjected to an SDS-PAGE electrophoresis using a 4-20% Criterion TGX gel (Bio-Rad). ..

    SDS Page:

    Article Title:
    Article Snippet: .. Same amounts of lysate, adjusted based on the total protein levels measured by Bradford assay, were subjected to an SDS-PAGE electrophoresis using a 4-20% Criterion TGX gel (Bio-Rad). ..

    Article Title: STING causes replication stress and nascent DNA degradation via SAMHD1
    Article Snippet: .. Equal protein amounts were separated by SDS-PAGE on a 4-15% Criterion TGX Gel (Bio-Rad) followed by transferring to a nitrocellulose membrane using the Trans-Blot Turbo system (Bio-Rad). ..

    Article Title:
    Article Snippet: .. The co-IP samples were visualized by performing SDS-PAGE electrophoresis using precast 4-20% Criterion TGX gel (Bio-Rad), staining the gel using Sypro Ruby (Invitrogen), and imaging the gel using ChemiDoc (Bio-Rad). .. The band intensity was quantified using ImageJ software by manually defining the rectangular area of each band, along with the background intensity from the neighboring area that was deducted from the band intensity.

    Article Title: Deletion of NLRP3 gene blocks traumatic brain injury induced abnormal immune response in 3xTg AD mice
    Article Snippet: The total protein concentration was determined using BCA method (Pierce, Rockford, IL). .. A 20μg of protein for each sample was boiled in Laemmli buffer (Bio-Rad, 1610747) and resolved using SDS-PAGE on a 4-20% Criterion TGX gel (Bio-Rad, 5671091). .. After SDS-PAGE, protein was transferred to a low-fluorescence PVDF (Bio-Rad) and blocked with either 5% BSA in tris-buffered saline with 0.1% Tween 20 (TBST) or 5% milk in TBST.

    Electrophoresis:

    Article Title:
    Article Snippet: .. Same amounts of lysate, adjusted based on the total protein levels measured by Bradford assay, were subjected to an SDS-PAGE electrophoresis using a 4-20% Criterion TGX gel (Bio-Rad). ..

    Article Title: Soluble expression of protein using peptide tag
    Article Snippet: .. For electrophoresis (SDS-PAGE) of protein, an electrophoresis tank (Criterion cell, BIO RAD) and Criterion TGX-gel (BIO RAD) were used. .. In the electrophoresis tank, an electrophoresis buffer (Tris/Glycine/SDS Buffer, BIO RAD) was placed, and 4 μl of the SDS-treated sample was applied to each well, followed by performing electrophoresis at a constant voltage of 200 V for 40 minutes.

    Article Title:
    Article Snippet: .. The co-IP samples were visualized by performing SDS-PAGE electrophoresis using precast 4-20% Criterion TGX gel (Bio-Rad), staining the gel using Sypro Ruby (Invitrogen), and imaging the gel using ChemiDoc (Bio-Rad). .. The band intensity was quantified using ImageJ software by manually defining the rectangular area of each band, along with the background intensity from the neighboring area that was deducted from the band intensity.

    Western Blot:

    Article Title: Yeast Smy2 and its human homologs GIGYF1 and -2 regulate Cdc48/VCP function during transcription stress
    Article Snippet: .. 50 mL 1x Sample Buffer was added and the sample boiled for 2 min 1% input and 20% sample were run on a 4–15% Criterion TGX gel (BioRad, 5671084) and normal Western blotting procedure followed. .. Proximity ligation assay Wild type or GIGYF1/2 DKO cells were plated in 8 well slides (PEZGS0816, Sigma).

    Nucleic Acid Electrophoresis:

    Article Title: Characterizing Mitochondrial Dysfunction Across Time in a Porcine model of Spinal Cord Injury
    Article Snippet: .. Mitochondrial samples were loaded onto a 4-20% Criterion TGX gel (BioRad, Cat. 5671095) and subjected to gel electrophoresis at 120V. .. Proteins were then transferred to a PVDF membrane (BioRad Midi format Trans-Blot Turbo Transfer Pack, Cat. 1704157) using the BioRad Trans-Blot Turbo Transfer System according to the preprogrammed Mixed MW setting (2.5A, up to 25V).

    Transferring:

    Article Title: STING causes replication stress and nascent DNA degradation via SAMHD1
    Article Snippet: .. Equal protein amounts were separated by SDS-PAGE on a 4-15% Criterion TGX Gel (Bio-Rad) followed by transferring to a nitrocellulose membrane using the Trans-Blot Turbo system (Bio-Rad). ..

    Staining:

    Article Title:
    Article Snippet: .. The co-IP samples were visualized by performing SDS-PAGE electrophoresis using precast 4-20% Criterion TGX gel (Bio-Rad), staining the gel using Sypro Ruby (Invitrogen), and imaging the gel using ChemiDoc (Bio-Rad). .. The band intensity was quantified using ImageJ software by manually defining the rectangular area of each band, along with the background intensity from the neighboring area that was deducted from the band intensity.

    Imaging:

    Article Title:
    Article Snippet: .. The co-IP samples were visualized by performing SDS-PAGE electrophoresis using precast 4-20% Criterion TGX gel (Bio-Rad), staining the gel using Sypro Ruby (Invitrogen), and imaging the gel using ChemiDoc (Bio-Rad). .. The band intensity was quantified using ImageJ software by manually defining the rectangular area of each band, along with the background intensity from the neighboring area that was deducted from the band intensity.



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    96
    Bio-Rad gradient polyacrylamide gel
    Relative HSP abundances in whole skeletal muscle homogenates from young adults and older adults pre and post HIT exercise. Representative Westen blots of (A) HSP72, HSP27, and αB-crystallin and (B) phosphorylated HSP27 Ser15 (pHSP27 Ser15) and pαB-crystallin Ser59 in whole muscle homogenates from the vastus lateralis of the same individuals. Calibration curves of mixed muscle homogenates are indicated and were used to determine the relative number of given proteins (see <t>Methods).</t> <t>Stain-free</t> gels are indicative of total protein loading, and molecular weights are indicated by markers collected under white light capture without moving the membrane between that and chemiluminescence detection. Relative abundances of (C) HSP72, (D) HSP27, (E) pHSP27 Ser15, (F) αB-crystallin, and (G) pαB-crystallin Ser59 from young (circle) and older adults Pre (square) and older adults Post (triangle) HIT exercise are shown relative to average Old (pre) on a given gel (data are presented as mean ± SD). Individuals indicated by the number of symbols ( n : 5–7), with the same color assigned to the same individual and consistent across all graphs. * p ≤ 0.05 indicates Brown-Forsye and Welch’s and post hoc analysis using Games-Horwell. HIT = high-intensity training; HSP = heat shock protein; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.
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    Relative HSP abundances in whole skeletal muscle homogenates from young adults and older adults pre and post HIT exercise. Representative Westen blots of (A) HSP72, HSP27, and αB-crystallin and (B) phosphorylated HSP27 Ser15 (pHSP27 Ser15) and pαB-crystallin Ser59 in whole muscle homogenates from the vastus lateralis of the same individuals. Calibration curves of mixed muscle homogenates are indicated and were used to determine the relative number of given proteins (see Methods). Stain-free gels are indicative of total protein loading, and molecular weights are indicated by markers collected under white light capture without moving the membrane between that and chemiluminescence detection. Relative abundances of (C) HSP72, (D) HSP27, (E) pHSP27 Ser15, (F) αB-crystallin, and (G) pαB-crystallin Ser59 from young (circle) and older adults Pre (square) and older adults Post (triangle) HIT exercise are shown relative to average Old (pre) on a given gel (data are presented as mean ± SD). Individuals indicated by the number of symbols ( n : 5–7), with the same color assigned to the same individual and consistent across all graphs. * p ≤ 0.05 indicates Brown-Forsye and Welch’s and post hoc analysis using Games-Horwell. HIT = high-intensity training; HSP = heat shock protein; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.

    Journal: Journal of Sport and Health Science

    Article Title: Exercise attenuates stress-related signaling as sensed by higher phosphorylation of small heat shock proteins in skeletal muscle from older individuals

    doi: 10.1016/j.jshs.2025.101111

    Figure Lengend Snippet: Relative HSP abundances in whole skeletal muscle homogenates from young adults and older adults pre and post HIT exercise. Representative Westen blots of (A) HSP72, HSP27, and αB-crystallin and (B) phosphorylated HSP27 Ser15 (pHSP27 Ser15) and pαB-crystallin Ser59 in whole muscle homogenates from the vastus lateralis of the same individuals. Calibration curves of mixed muscle homogenates are indicated and were used to determine the relative number of given proteins (see Methods). Stain-free gels are indicative of total protein loading, and molecular weights are indicated by markers collected under white light capture without moving the membrane between that and chemiluminescence detection. Relative abundances of (C) HSP72, (D) HSP27, (E) pHSP27 Ser15, (F) αB-crystallin, and (G) pαB-crystallin Ser59 from young (circle) and older adults Pre (square) and older adults Post (triangle) HIT exercise are shown relative to average Old (pre) on a given gel (data are presented as mean ± SD). Individuals indicated by the number of symbols ( n : 5–7), with the same color assigned to the same individual and consistent across all graphs. * p ≤ 0.05 indicates Brown-Forsye and Welch’s and post hoc analysis using Games-Horwell. HIT = high-intensity training; HSP = heat shock protein; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.

    Article Snippet: Total protein was separated on 10% or 4%–15% Criterion TGX stain-free gels (Bio-Rad Laboratories) and run for 45 min at 200 V. Using a wet transfer protocol, protein was transferred to nitrocellulose membranes at 100 V for 30 min. Membranes were incubated in Miser TM solution (ThermoFisher Scientific) and blocked in 5% skim milk powder in tris-buffered saline-tween (TBST).

    Techniques: Staining, Membrane

    HSP abundances in type I and II skeletal muscle fibers from young and older adults. (A, C, and F) The MHC isoform present was determined in individual muscle fiber segments from the vastus lateralis and, following pooling into type I and type II groups from a given biopsy, were analyzed by Westen blotting. Westen blots of (A) HSP72, (C) HSP27 and pHSP27 Ser15, (F) αB-crystallin and pαB-crystallin Ser59, with MHC isoforms in groups of fibers. Stain-free gels are indicative of total protein loading, and molecular weights are indicated by markers collected under white light capture without moving the membrane between that and chemiluminescence detection. Calibration curves of mixed muscle homogenates are indicated. Relative protein abundances of (B) HSP72, (D) HSP27, (E) pHSP27 Ser15, (G) αB-crystallin, and (H) pαB-crystallin Ser59 in fibers from young (circle) and older adults (square) type I fibers (no outline) and type II fibers (outline). All fibers are expressed relative to the average older adult’s type I fibers. The same color is assigned to the same individual and is consistent with (data are presented as mean ± SD). * p < 0.05 and ** p < 0.01, mixed effect model Univariant using either Tukey’s or Games-Horwell’s multiple comparison test (see Methods). HIT = high-intensity training; HSP = heat shock protein; MHC = myosin heavy chain; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.

    Journal: Journal of Sport and Health Science

    Article Title: Exercise attenuates stress-related signaling as sensed by higher phosphorylation of small heat shock proteins in skeletal muscle from older individuals

    doi: 10.1016/j.jshs.2025.101111

    Figure Lengend Snippet: HSP abundances in type I and II skeletal muscle fibers from young and older adults. (A, C, and F) The MHC isoform present was determined in individual muscle fiber segments from the vastus lateralis and, following pooling into type I and type II groups from a given biopsy, were analyzed by Westen blotting. Westen blots of (A) HSP72, (C) HSP27 and pHSP27 Ser15, (F) αB-crystallin and pαB-crystallin Ser59, with MHC isoforms in groups of fibers. Stain-free gels are indicative of total protein loading, and molecular weights are indicated by markers collected under white light capture without moving the membrane between that and chemiluminescence detection. Calibration curves of mixed muscle homogenates are indicated. Relative protein abundances of (B) HSP72, (D) HSP27, (E) pHSP27 Ser15, (G) αB-crystallin, and (H) pαB-crystallin Ser59 in fibers from young (circle) and older adults (square) type I fibers (no outline) and type II fibers (outline). All fibers are expressed relative to the average older adult’s type I fibers. The same color is assigned to the same individual and is consistent with (data are presented as mean ± SD). * p < 0.05 and ** p < 0.01, mixed effect model Univariant using either Tukey’s or Games-Horwell’s multiple comparison test (see Methods). HIT = high-intensity training; HSP = heat shock protein; MHC = myosin heavy chain; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.

    Article Snippet: Total protein was separated on 10% or 4%–15% Criterion TGX stain-free gels (Bio-Rad Laboratories) and run for 45 min at 200 V. Using a wet transfer protocol, protein was transferred to nitrocellulose membranes at 100 V for 30 min. Membranes were incubated in Miser TM solution (ThermoFisher Scientific) and blocked in 5% skim milk powder in tris-buffered saline-tween (TBST).

    Techniques: Staining, Membrane, Comparison