Gapdh mouse monoclonal antibody

Manufactured by Beyotime

Sourced in United States, China

The GAPDH Mouse Monoclonal Antibody is a laboratory reagent used to detect and quantify the presence of the GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) protein in biological samples. This antibody is specific to the GAPDH protein and can be used in various immunoassay techniques, such as Western blotting, immunohistochemistry, and ELISA, to analyze the expression levels of GAPDH in cells or tissues.

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Lab products found in correlation

Hrp labeled goat anti mouse igg h l, by Beyotime (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Protease inhibitor pmsf, by Beyotime (1 mentions) Sds page gel, by Beyotime (1 mentions) Immobilon fl pvdf, by Merck Group (1 mentions) Beyoecl plus reagent, by Beyotime (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Bromophenol blue, by Bio-Rad (1 mentions) Aspirin, by Merck Group (1 mentions) Coomassie brilliant blue g 250, by Bio-Rad (1 mentions) Tris base, by Bio-Rad (1 mentions) Tetramethylethylenediamine, by Bio-Rad (1 mentions) Human ox ldl bt 910, by Thermo Fisher Scientific (1 mentions) Itraq kit, by Thermo Fisher Scientific (1 mentions) Sirna reagent, by Santa Cruz Biotechnology (1 mentions) Sequence grade modified trypsin, by Promega (1 mentions) 2 d quant kit, by GE Healthcare (1 mentions) Nitrocellulose membrane, by Bio-Rad (1 mentions) High performance liquid chromatography system, by Shimadzu (1 mentions)

6 protocols using gapdh mouse monoclonal antibody

Chicken Cell Lines for IBV Studies

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Chicken macrophage cell line HD11 was presented by Professor Jiao Xin’an from Yangzhou University and maintained in Dulbecco’s modified Eagle’s medium (DMEM) (HyClone), supplemented with 10% fetal bovine serum (FBS) (Gibco) and 1% penicillin-streptomycin (DMEM growth medium). Chicken fibroblast cell line DF-1 was from our laboratory and maintained in DMEM (HyClone), supplemented with 10% FBS (Gibco) and 1% penicillin-streptomycin (DMEM growth medium).
Both IBV Beaudette strain and IBV M41 strain (standard strain) were from our laboratory. For WB and immunofluorescence assays, the Flag-tag mouse monoclonal antibody (Beyotime Biotechnology), HRP-labeled goat anti-mouse IgG(H + L) (Beyotime Biotechnology), GAPDH Mouse Monoclonal Antibody (Beyotime Biotechnology), and Alexa Fluor 350 labeled goat anti-rabbit IgG (H + L) (Beyotime Biotechnology) were used as primary and second antibodies, respectively.

Wang K., Cui P., Ni R., Gong H., Li H., Yan W., Fu X., Chen L., Lei C., Wang H, & Yang X. (2022). Chicken-Derived Pattern Recognition Receptor chLGP2 Inhibits the Replication and Proliferation of Infectious Bronchitis Virus. Frontiers in Microbiology, 12, 810215.

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Comprehensive Protein Analysis Pipeline

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RIPA lysis buffer (Beyotime, China) and protease inhibitor PMSF (Beyotime, China) were used to lyse the cells, and protein concentration was evaluated by utilizing Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, USA). The protein samples were added into SDS-PAGE gels (Beyotime, China) in the electrophoresis apparatus for indicated period, followed by being transferred to Immobilon-FL PVDF (Merck, Germany). Next, PVDF was incubated with 5% non-fat milk for 90 min at room temperature, followed by been incubated with primary antibodies (A20 Rabbit mAb, Cell Signaling Technology, USA; Rabbit monoclonal to Collagen I, Abcam, USA; Rabbit monoclonal to Collagen III, Abcam, USA; Rabbit monoclonal to Fibronectin, Abcam, USA; RIP Rabbit mAb, Cell Signaling Technology, USA; GAPDH Mouse Monoclonal Antibody, Beyotime, China) overnight at 4 °C. After incubation with secondary antibodies (Beyotime, China), PVDF was detected with BeyoECL Plus reagent (Beyotime, China).

Kuai Q, & Jian X. (2022). Inhibition of miR-23b-3p Ameliorates Scar-Like Phenotypes of Keloid Fibroblasts by Facilitating A20 Expression. Clinical, Cosmetic and Investigational Dermatology, 15, 1549-1559.

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HSYA Isolation and Analysis in C. tinctorius

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HSYA was isolated and purified from the aqueous extract of C. tinctorius L. by macroporous resin-gel column chromatography, as described previously^{14 (link)}. The molecular weight of HSYA is 612. HSYA was analysed using a high-performance liquid chromatography system (Shimadzu, Kyoto, Japan). iTRAQ kits were purchased from Applied Biosystems (Waltham, MA, USA), the 2-D Quant Kit from GE Healthcare (Pittsburgh, PA, USA), sequence-grade modified trypsin from Promega (Madison, WI, USA), and human ox-LDL (BT-910) from Alfa Aesar (Heysham, Lancashire, UK). Aspirin was the product of Sigma-Aldrich (St. Louis, MO, USA). Bromophenol blue, tetramethylethylenediamine, Coomassie brilliant blue G-250, Bis, low-molecular weight marker, nitrocellulose membrane, Tris-base, and the Bradford method protein assay kit were purchased from Bio-Rad (Hercules, CA, USA). VDAC-2 polyclonal antibody was purchased from Cell Signaling Technology (Danvers, MA, USA), and GAPDH mouse monoclonal antibody was from Beyotime Biotechnology (Jiangsu, China). Other polyclonal and monoclonal antibodies and siRNA reagents were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), with the secondary antibodies obtained from LI-COR Biosciences (Lincoln, NE, USA). NO and MDA assay kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Ye F., Wang J., Meng W., Qian J, & Jin M. (2017). Proteomic investigation of effects of hydroxysafflor yellow A in oxidized low-density lipoprotein-induced endothelial injury. Scientific Reports, 7, 17981.

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Western Blot Analysis of Apoptosis Proteins

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Cells were directly lysed using a home-made loading buffer (12 mM Tris-HCl (pH6.8), 5% glycerol, 0.4% SDS, 2.88 mM β-mercaptoethanol, 0.02% bromophenol blue). Protein extracts were boiled for 10 min and then separated on SDS–PAGE gels. Protein amounts were adjusted according to the housekeeping gene GAPDH.
Antibodies used for Western blots were as follows: SERPINB2 rabbit polyclonal antibody (ABclonal, A15297, Wuhan, China), GAPDH mouse monoclonal antibody (Beyotime Biotechnology, AF0006), Caspase-3 rabbit polyclonal antibody (Beyotime Biotechnology, AF0081). Secondary antibodies were HRP-conjugated goat anti-mouse IgG (Beyotime Biotechnology, A0216) and peroxidase-conjugated goat anti-rabbit IgG (Beyotime Biotechnology, A0208).

Chen Z., Wei Y., Zheng Y., Zhu H., Teng Q., Lin X., Wu F, & Zhou F. (2022). SERPINB2, an Early Responsive Gene to Epigallocatechin Gallate, Inhibits Migration and Promotes Apoptosis in Esophageal Cancer Cells. Cells, 11(23), 3852.

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Protein Quantification and Western Blotting Protocol

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Western blotting was conducted as previously described [8 (link)]. Similarly, we obtained the supernatant of the sample and extracted proteins using the radio immunoprecipitation assay (RIPA) buffer containing 1 mmol/L PMSF (Beyotime, Shanghai, China), and the protein concentration was quantified using a BCA assay kit (Beyotime, Shanghai, China). During the process of electrophoresis, the proteins with different sizes were separated in SDS-PAGE. Then, the proteins on the gel were transferred onto a polyvinylidene-difluoride (PVDF) membrane (Beyotime, Shanghai, China) for blotting. Antibodies were acquired from Beyotime Biotechnology, Shanghai, China, which included glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mouse monoclonal antibody (catalog number: AG019), caspase-1rabbit polyclonal antibody (catalog number: AF1681), Nrf2 rabbit polyclonal antibody (catalog number: AF7623), NF-κB p65 rabbit polyclonal antibody (catalog number: AF5875), HO-1rabbit polyclonal antibody (catalog number: AF1333), NLRP3 rabbit monoclonal antibody (catalog number: AF2155), HRP-labeled goat anti-rabbit IgG (H + L) (catalog number: A0208) and HRP-labeled goat anti-mouse IgG (H + L) (catalog number: A0216). Original Western Blot figures in Figure S2.

Yang H., Wang Y., Liu M., Liu X., Jiao Y., Jin S., Shan A, & Feng X. (2021). Effects of Dietary Resveratrol Supplementation on Growth Performance and Anti-Inflammatory Ability in Ducks (Anas platyrhynchos) through the Nrf2/HO-1 and TLR4/NF-κB Signaling Pathways. Animals : an Open Access Journal from MDPI, 11(12), 3588.

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Western Blot Protein Quantification Protocol

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Cells were lysed by RIPA buffer to obtain total protein lysates and then separated by 10% SDS-polyacrylamide gel electrophoresis (SDS-PAGE). After transferred to 0.22 μm NC membranes (Sigma), protein lysates were incubated with specific antibodies. The ECL chromogenic substrate was used to quantify by densitometry (Quantity One software; Bio-Rad). The following primary antibodies were used: Cyclin D3 (Proteintech, no. 26755-1-AP), CDK4 (MXB, no. EP180), OAS1 (D1W3A) Rabbit mAb (Cell Signaling Technology, no. 14498), Cleaved Casepase-3 (Asp175) Antibody (Cell Signaling Technology, no. 9661, PARP (46D11) Rabbit mAb (Cell Signaling Technology, no. 9532), Casepase-3 Antibody (Cell Signaling Technology, no. 9662), Cleaved PARP (Asp214) (D64E10) XP^® Rabbit mAb (Cell Signaling Technology, no. 5625), β-Actin Mouse Monoclonal Antibody (Beyotime, no. AF5001), GAPDH Mouse Monoclonal Antibody (Beyotime, no AF5009), Anti-rabbit IgG, HRP-linked Antibody (Cell Signaling Technology, no. 7074S), Anti-mouse IgG, HRP-linked Antibody (Cell Signaling Technology, no. 7076S). The β-actin antibody and GAPDH antibody were used as control.

Lu D., Di S., Zhuo S., Zhou L., Bai R., Ma T., Zou Z., Chen C., Sun M., Tang J, & Zhang Z. (2021). The long noncoding RNA TINCR promotes breast cancer cell proliferation and migration by regulating OAS1. Cell Death Discovery, 7, 41.

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