Mouse anti gapdh

Manufactured by Biotrend

Sourced in Germany, United States

Mouse anti-GAPDH is a primary antibody that specifically binds to the GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) protein, which is a crucial enzyme involved in the glycolytic pathway. It can be used for the detection and quantification of GAPDH in various research applications.

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Anti-GAPDH (2 citations)

4 protocols using mouse anti gapdh

Comprehensive Western Blot Analysis of Cell Death Regulators

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Western blot analysis was performed as described previously [29] (link), using the following antibodies: rabbit anti-MLKL, rabbit anti-RIPK3, rabbit anti-phospho MLKL, rabbit anti-phospho RIKP1, rabbit anti-phospho RIPK3, rabbit anti-TNFR1, rabbit anti-cIAP2 (all from Cell Signaling), mouse anti-RIPK1 (BD Bioscience), mouse anti-GAPDH (BioTrend), rabbit anti-TRAIL-R1 (Abcam), rabbit anti-TRAIL-R2 (Millipore), rabbit anti-FasR, mouse anti-cIAP1 (all from Santa Cruz), mouse anti-Caspase-8 (Enzo). For detection, secondary antibodies goat anti-mouse IgG and goat anti-rabbit IgG (Abcam) conjugated to horseradish peroxidase and enhanced chemiluminescence were used (Amersham Bioscience).

Koch A., Jeiler B., Roedig J., van Wijk S.J., Dolgikh N, & Fulda S. (2021). Smac mimetics and TRAIL cooperate to induce MLKL-dependent necroptosis in Burkitt's lymphoma cell lines. Neoplasia (New York, N.Y.), 23(5), 539-550.

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Western Blot Analysis of Apoptosis Regulators

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Protein extraction was done with TritonX-containing lysis buffer and protein content was determined using BCA assay (Pierce^TM BCA protein assay, Thermo Fisher Scientific). SDS–PAGE was carried out followed by semidry blotting. After blocking the membrane for 1 h with 5% milk, proteins were detected using the following antibodies: rabbit anti-MCL-1 (Enzo, ADI-AAP-240F), mouse anti-BCL-2 (Dako (Agilent), M088701-2), rabbit anti-BCL-x_L (CellSignaling, 2762 S), mouse anti-NOXA (Enzo, ALX-804-408), mouse anti-BAX (BD Bioscience, 610983), rabbit anti-BAK (Upstate/ Merck, 06–536), rabbit anti-BIM (CellSignaling, 3183 S) and rabbit anti-caspase-3 (CellSignaling, 9662 S), mouse anti-β-Actin (Sigma, A5441), mouse anti-GAPDH (BioTrend (Hy Test Ltd), 5G4-6C5) or mouse anti-Vinculin (Sigma/Merck, V9131-100UL). Quantification of protein expression was performed using ImageJ 3.1 software.

Ewald L., Dittmann J., Vogler M, & Fulda S. (2019). Side-by-side comparison of BH3-mimetics identifies MCL-1 as a key therapeutic target in AML. Cell Death & Disease, 10(12), 917.

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Immunoblotting of Cell Signaling Proteins

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Immunoblotting was performed using the following antibodies: mouse anti-cIAP1 (Santa Cruz, Heidelberg, Germany; 271418), rabbit anti-cIAP2 (Cell Signaling Technologies, Leiden, The Netherlands; 3130), rabbit anti-survivin (R&D; AF886), mouse anti-XIAP (BD, Franklin Lakes, NJ, USA; 610716), rabbit anti-IκBα (Cell Signaling Technologies; 9242), mouse anti-p-IκBα (Cell Signaling Technologies; 6249L), mouse anit-p65 (Santa Cruz; sc-8008), rabbit anti-p-p65 (Cell Signaling Technologies; 3033S), rabbit anti-NIK (Cell Signaling Technologies; 4994), mouse anti-p100 (Merck-Millipore, Darmstadt, Germany; 05-361), mouse anti-GAPDH (BioTrend, Köln, Germany; NB-29-00852), mouse anti-vinculin (Sigma/Merck; V9131), mouse anti-caspase-8 (Enzo; ADI-AAM-118-E), rabbit anti-caspase-9 (Cell Signaling Technologies; 9502S), rabbit anti-DR5 (Merck-Millipore; AB16942) and mouse anti-PARP-1 (Cell Signaling Technologies; 9546S). The following secondary horseradish peroxidase-conjugated antibodies were used: goat anti-mouse (Abcam, Berlin, Germany; ab6789) and goat anti-rabbit (Abcam; ab6721). The quantification of Western blot and the area determination of the region of interest were performed using ImageJ/FIJI [28 (link)]. Subsequent ratio calculations were performed using Microsoft Excel.

Särchen V., Reindl L.M., Wiedemann S., Shanmugalingam S., Bukur T., Becker J., Suchan M., Ullrich E, & Vogler M. (2023). Characterization of BV6-Induced Sensitization to the NK Cell Killing of Pediatric Rhabdomyosarcoma Spheroids. Cells, 12(6), 906.

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Western Blot Analysis of Autophagy Proteins

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Western blot analysis was performed as described previously using RIPA buffer (50 mM Tris-HCl, pH 8, 1% Triton-X, 0.5% sodium deoxycholate, 150 mM sodium chloride and 2 mM magnesium chloride) supplemented with Pierce Nuclease (Thermo Fisher, Waltham, MA, USA)^{83 (link)}. The following antibodies were used: monoclonal rabbit anti-ATG7, rabbit anti-ATG5 (Cell signalling, Danvers, Massachusetts, USA), mouse anti-GAPDH (Biotrend, Cologne, Germany), mouse anti-vinculin (Sigma, Germany), rabbit anti-LC3B (Thermo Fisher, Waltham, MA, USA), mouse anti-Mitofusin-2, mouse anti-COXIV (Abcam, Cambridge, UK) and rabbit anti-RFP (ChromoTek, Martinsried, Germany). Goat anti-mouse and goat anti-rabbit conjugated to horseradish peroxidase (Santa Cruz Biotechnology, Santa Cruz, CA, USA) as well as enhanced chemiluminescence (Amersham Biosciences, Freiburg, Germany) were used for detection. Representative blots of at least two independent experiments are shown.

Kinzler M.N., Zielke S., Kardo S., Meyer N., Kögel D., van Wijk S.J, & Fulda S. (2020). STF-62247 and pimozide induce autophagy and autophagic cell death in mouse embryonic fibroblasts. Scientific Reports, 10, 687.

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