Anti gapdh mouse mab

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-GAPDH mouse MAb is a mouse monoclonal antibody that recognizes Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), a key enzyme involved in glycolysis.

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Lab products found in correlation

Horseradish peroxidase hrp conjugated goat anti mouse or anti rabbit igg, by Santa Cruz Biotechnology (4 mentions) Anti flag m2 rabbit mab, by Cell Signaling Technology (4 mentions) Anti α tubulin mouse mab, by Santa Cruz Biotechnology (4 mentions) Anti his mouse mab, by Beyotime (3 mentions) Anti mouse immunoglobulin g igg, by Beyotime (3 mentions) Anti phospho c met y1356 rabbit polyclonal antibody pab, by Abcam (2 mentions) Anti c met rabbit mab, by Cell Signaling Technology (2 mentions) Anti cd82 rabbit pab, by Santa Cruz Biotechnology (2 mentions) Anti b fgf goat pab, by Santa Cruz Biotechnology (2 mentions) Anti gfp rabbit antibody, by Beyotime (2 mentions) Pf 2341066, by Selleck Chemicals (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Effectence transfection reagent, by Qiagen (2 mentions) Pvdf membrane, by Merck Group (2 mentions) Ab126090, by Abcam (1 mentions) Chloroquine, by Merck Group (1 mentions) Mouse anti myc mab, by Santa Cruz Biotechnology (1 mentions) Cycloheximide (chx), by Merck Group (1 mentions) Mouse anti gm130 mab, by BD (1 mentions) Mouse anti gfp mab, by Santa Cruz Biotechnology (1 mentions)

Close spelling variants of this product

Anti-GAPDH mAb GAPDH mAb Mouse anti-GAPDH GAPDH (mouse, Anti-GAPDH GAPDH

10 protocols using anti gapdh mouse mab

Western Blot Analysis of Protein Expression

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Cells
were seeded in 6-well plates (1 × 10⁶ cells per well).
After overnight incubation, the cells were treated with indicated
concentrations for specific timepoints. The harvested cells were spinned
down, washed with PBS, and lysed with 1× LDS loading buffer followed
by sonication and heated at 70 °C. The prepared samples were
loaded on NuPAGE 4–12% Bis–Tris protein gels and transferred
to the nitrocellulose membrane. After blocking the membrane with LI-COR
TBS blocking buffer and incubation in primary antibody overnight,
the corresponding protein signals were detected using IRDye secondary
antibodies and an Odyssey imaging system. Image Studio Lite Ver 5.2
was used for blot quantification. Rabbit anti-human eRF3/GSPT1 pAb
(ab126090, Abcam), mouse anti-human Ikaros mAb (sc-398265, Santa Cruz
Biotechnology), mouse anti-GAPDH mAb (sc-47724, Santa Cruz Biotechnology)
were used as primary antibodies. The IRDye 800CW Goat anti-Mouse IgG
Secondary Antibody and IRDye 680RD Goat anti-Rabbit IgG Secondary
Antibody were used as secondary antibodies.

Nishiguchi G., Keramatnia F., Min J., Chang Y., Jonchere B., Das S., Actis M., Price J., Chepyala D., Young B., McGowan K., Slavish P.J., Mayasundari A., Jarusiewicz J.A., Yang L., Li Y., Fu X., Garrett S.H., Papizan J.B., Kodali K., Peng J., Pruett Miller S.M., Roussel M.F., Mullighan C., Fischer M, & Rankovic Z. (2021). Identification of Potent, Selective, and Orally Bioavailable Small-Molecule GSPT1/2 Degraders from a Focused Library of Cereblon Modulators. Journal of Medicinal Chemistry, 64(11), 7296-7311.

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Immunofluorescence Staining Protocol

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Mouse anti-EEA1 monoclonal antibody (mAb) (#610456; 1:500 for immunofluorescence or IF) and mouse anti-GM130 mAb (#610823; 1:500 for IF) were from BD Biosciences. Mouse anti-Lamp1 mAb (H4A3; 1:500 for IF) was from Developmental Studies Hybridoma Bank. Mouse anti-Myc mAb (#sc-40; 1:200 for IF), mouse anti-GAPDH mAb (#sc-47724; 1:1000 for Western blot) and mouse anti-GFP mAb (#sc-25778; 1:1000 for Western blot) were from Santa Cruz. Rabbit anti-GFP (Mahajan et al., 2019) and anti-mCherry (Sun et al., 2020) polyclonal sera were previously described. Horseradish peroxidaseconjugated goat secondary antibodies were from Bio-Rad. Alexa Fluor-conjugated goat antibodies against mouse or rabbit IgG (1:500 for IF) were from Thermo Fisher Scientific. CHX (working concentration: 10 µg/ml) and chloroquine (working concentration: 50 µM) were from Sigma Aldrich. Bafilomycin A1 (working concentration: 100 nM) was from Chemscene.

Sun X., Chen B., Song Z., & Lü L. (2021). A quantitative study of the Golgi retention of glycosyltransferases.

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Inhibition of KSHV-mediated STAT3 Signaling

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Anti-KSHV LANA rat monoclonal antibody (MAb) was purchased from Advanced Biotechnologies Inc.(Columbia, MD, USA) [81 (link)]. Anti-phospho-STAT3 (Y705) rabbit MAb, anti-STAT3 mouse polyclonal antibody (PAb), and anti-Flag M2 rabbit MAb were obtained from Cell Signaling Technologies (Beverly, MA, USA). Anti-SH3BGR mouse MAb, anti-GAPDH mouse MAb, anti-α-Tubulin mouse MAb, and horseradish peroxidase (HRP)-conjugated goat anti-mouse or anti-rabbit IgG were all purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-His mouse MAb, anti-mouse immunoglobulin G (IgG) and anti-GST mouse MAb were from Beyotime Institute of Biotechnology (Nantong, Jiangsu, China). Western blotting analysis was performed as described previously [84 (link), 85 (link)]. AG490, a JAK2 inhibitor, was purchased from Selleck Chemicals (Shanghai, China).

Li W., Yan Q., Ding X., Shen C., Hu M., Zhu Y., Qin D., Lu H., Krueger B.J., Renne R., Gao S.J, & Lu C. (2016). The SH3BGR/STAT3 Pathway Regulates Cell Migration and Angiogenesis Induced by a Gammaherpesvirus MicroRNA. PLoS Pathogens, 12(4), e1005605.

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SAMHD1 Protein Phosphorylation Analysis

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Samples were processed in RIPA buffer containing 1 μM DTT, 10 μM PMSF, 10 μl/ml phosphatase inhibitor (Sigma) and 10 μl/ml protease inhibitor (Sigma). The cells were sonicated with 3×, 5 second pulses, to ensure complete lysis. Cellular debris was removed by 15 K rpm centrifugation for 10 min. Supernatants were stored at −80°C before use. Cell lysates were resolved on a 8% SDS-PAGE gel. Proteins were transferred to nitrocellulose membrane and detected as described in the figure legends using the following antibodies: rabbit anti-SAMHD1 mAb (Abcam), anti-GAPDH mouse mAb (Santa Cruz). Anti-mouse and anti-rabbit secondary HRP antibodies were purchased from GE HealthScience. HRP was detected using chemiluminescent reagents (Pierce) following the manufacturers instructions. Images were captured using BioRad ChemiDoc Imager. Anti-pSAMHD1 T592 antibody was obtained from Dr. Diaz-Griffero.

Hollenbaugh J.A., Tao S., Lenzi G.M., Ryu S., Kim D.H., Diaz-Griffero F., Schinazi R.F, & Kim B. (2014). dNTP pool modulation dynamics by SAMHD1 protein in monocyte-derived macrophages. Retrovirology, 11, 63.

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Western Blot Analysis of KSHV LANA

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Anti-KSHV LANA ORF73 rat monoclonal antibody (MAb) was purchased from Advanced Biotechnologies Inc. (Columbia, MD, USA) [78 (link)]. Anti-phospho-AKT (Ser473) rabbit MAb, anti-AKT rabbit polyclonal antibody (PAb), anti-Flag M2 rabbit MAb, anti-His rabbit MAb and anti-HA rabbit PAb were obtained from Cell Signaling Technologies (Beverly, MA, USA). Anti-GRK2 mouse MAb, anti-CXCR2 rabbit PAb, anti-GAPDH mouse MAb, anti-α-Tubulin mouse MAb, and horseradish peroxidase (HRP)-conjugated goat anti-mouse or anti-rabbit IgG were all purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-GFP mouse MAb was from Beyotime Institute of Biotechnology (Nantong, Jiangsu, China). Western blotting analysis was performed as described previously [83 (link)]. MK-2206, an AKT inhibitor, was purchased from Selleck Chemicals (Shanghai, China).

Hu M., Wang C., Li W., Lu W., Bai Z., Qin D., Yan Q., Zhu J., Krueger B.J., Renne R., Gao S.J, & Lu C. (2015). A KSHV microRNA Directly Targets G Protein-Coupled Receptor Kinase 2 to Promote the Migration and Invasion of Endothelial Cells by Inducing CXCR2 and Activating AKT Signaling. PLoS Pathogens, 11(9), e1005171.

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Western Blot Analysis of KIAA1199 and TMEM2

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Total cell extract was prepared with 1 × RIPA buffer [50 mmol/L-Tris-HCl buffer (pH 7.6), 150 mmol/L-NaCl, 1 %-Nonidet P40 substitute (w/v), 0.5 %-sodium deoxycholate (w/v), protease inhibitor cocktail, 0.1 %-SDS (w/v)] (FUJIFILM Wako Pure Chemical). The samples were separated on 10% SuperSep Ace gels (Wako) and transferred to PVDF membranes (Merck). Nonspecific binding was blocked at room temperature (15–25 °C) for 15 min with Blocking One-P (Nacalai Tesque, Kyoto, Japan). The membranes were incubated overnight at 4 °C with the following antibodies: anti-KIAA1199 (HYBID) rabbit polyclonal antibody (pAb) (1:1000) (Sigma Aldrich, MO, USA), anti-TMEM2 rabbit pAb (1:500) (Sigma Aldrich), anti-FLAG M2 mouse mAb (1:2000) (Sigma Aldrich) and anti-GAPDH mouse mAb (1:2000) (Santa Cruz Biotechnology). The primary antibodies were detected using HRP-conjugated secondary antibodies (GE Healthcare). Bands were detected via ImmunoStar LD (FUJIFILM Wako Pure Chemical) using an Amersham Imager 680 (GE Healthcare Bioscience, NJ, USA).

Sato S., Miyazaki M., Fukuda S., Mizutani Y., Mizukami Y., Higashiyama S, & Inoue S. (2023). Human TMEM2 is not a catalytic hyaluronidase, but a regulator of hyaluronan metabolism via HYBID (KIAA1199/CEMIP) and HAS2 expression. The Journal of Biological Chemistry, 299(6), 104826.

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Western Blot Analysis of Key Signaling Proteins

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Proteins were separated by 8% and 10% SDS polyacrylamide gels and electrophoretically transferred onto polyvinylidene fluoride membranes (Millipore, Billerica, MA, USA). The membranes were blocked overnight with 5% non-fat dried milk and incubated overnight with antibodies. Anti-Kidins220 rabbit polyclonal antibody (PAb) (Cat No. ab97345) was purchased from Abcam Inc (Abcam, Cambridge, UK). Anti-PI3K rabbit monoclonal antibody (MAb) (Cat No. ♯4257), anti-phospho-PI3K rabbit PAb (Cat No. ♯4228), anti-AKT rabbit PAb (Cat No. ♯9272) and anti-phospho-AKT (Cat No. ♯4060) were all purchased from Cell Signaling Technologies (Beverly, MA, USA). Anti-VEGF rabbit PAb (Cat No. sc-152), anti-GAPDH mouse MAb (Cat No. sc-47724), anti-a-Tubulin mouse MAb (Cat No. sc-23948) and horseradish peroxidase (HRP) conjugated goat anti-rabbit (Cat No. sc-2004) or anti-mouse (Cat No. sc-2005) secondary antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The signals were detected by enhanced chemiluminescence (Millipore).

Wang Y., Shao N., Mao X., Zhu M., Fan W., Shen Z., Xiao R., Wang C., Bao W., Xu X., Yang C., Dong J., Yu D., Wu Y., Zhu C., Wen L., Lu X., Lu Y.J, & Feng N. (2016). MiR-4638-5p inhibits castration resistance of prostate cancer through repressing Kidins220 expression and PI3K/AKT pathway activity. Oncotarget, 7(30), 47444-47464.

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Western Blot Analysis of KSHV LANA and c-Met

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An anti-KSHV LANA rat monoclonal antibody (MAb) was purchased from Advanced Biotechnologies Inc. (Columbia, MD, USA)^{89 (link)}. The anti-phospho-c-Met (Y1356) rabbit polyclonal antibody (PAb) was obtained from Abcam (Cambridge, MA, USA). The anti-c-Met rabbit MAb and anti-Flag M2 rabbit MAb were obtained from Cell Signaling Technologies (Beverly, MA, USA). The anti-CD82 rabbit PAb, anti-b-FGF goat PAb, anti-GAPDH mouse MAb, anti-α-Tubulin mouse MAb, and horseradish peroxidase (HRP)-conjugated goat anti-mouse or anti-rabbit IgG were all purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The anti-His mouse MAb, anti-GFP rabbit antibody, and anti-mouse immunoglobulin G (IgG) were from Beyotime Institute of Biotechnology (Nantong, Jiangsu, China). Western-blotting analysis was performed as previously described^{90 (link), 91 (link)}. Transfection of HUVECs was performed with the Effectence transfection reagent (Qiagen, Valencia, CA, USA) while transfections in other cells were performed with Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) following the manufacturer’s instructions. PF-2341066, a c-Met inhibitor, was purchased from Selleck Chemicals (Shanghai, China).

Li W., Hu M., Wang C., Lu H., Chen F., Xu J., Shang Y., Wang F., Qin J., Yan Q., Krueger B.J., Renne R., Gao S.J, & Lu C. (2017). A Viral MicroRNA Down-regulates Metastasis Suppressor CD82 and Induces Cell Invasion and Angiogenesis by Activating the c-Met Signaling. Oncogene, 36(38), 5407-5420.

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Western Blot Analysis of KSHV LANA and c-Met

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Western Blot Analysis of STAT and Inflammasome Proteins

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Cell lysis was carried out in a lysis buffer containing 50 mM Tris (pH 7.5), 1% (v/v) Triton X-100, 150 mM NaCl, 10% (v/v) glycerol, 1 mM EDTA, and a protease inhibitor cocktail. The resulting lysates were resolved by 10% SDS-PAGE, and then were transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore, USA). These samples were incubated with mouse polyclonal antibodies (pAbs) and horseradish peroxidase-conjugated goat anti-mouse secondary antibodies (1:10000, Santa Cruz Biotechnology) overnight at 4 °C. The pAbs were raised against STAT-1, phospho-STAT-1, STAT-3, phospho-STAT-3, proIL-1β, pro caspase-1, and caspase-1 (1:1000, Cell Signaling Technology). Then, we used anti-GAPDH mouse mAb (1:1000, Santa Cruz) for normalization. Finally, an ECL Western blotting analysis system (Cell Signaling Technology) was used for visualization.

Sun Y., Ma J., Li D., Li P., Zhou X., Li Y., He Z., Qin L., Liang L, & Luo X. (2019). Interleukin-10 inhibits interleukin-1β production and inflammasome activation of microglia in epileptic seizures. Journal of Neuroinflammation, 16, 66.

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