Anti gapdh antibody 14c10

Manufactured by Cell Signaling Technology

The Anti-GAPDH antibody (14C10) is a primary antibody used to detect the GAPDH protein, which is a widely expressed enzyme involved in glycolysis. This antibody can be used to identify and quantify GAPDH levels in various sample types through techniques such as Western blotting.

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Lab products found in correlation

Peroxidase conjugated secondary antibody, by GE Healthcare (2 mentions) Skim milk, by Nacalai Tesque (2 mentions) Immobilon p pvdf membrane, by Merck Group (2 mentions) Anti mcm2 antibody d7g11, by Cell Signaling Technology (2 mentions) Anti cdc6 antibody sc 9964, by Santa Cruz Biotechnology (2 mentions) Anti cdt1 d10f11, by Cell Signaling Technology (2 mentions) Anti orc2 antibody 3g6, by Santa Cruz Biotechnology (2 mentions) Anti cdc45 antibody d7g6, by Cell Signaling Technology (2 mentions) Anti histone h3 antibody, by Santa Cruz Biotechnology (1 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions) Colloidal blue staining kit, by Thermo Fisher Scientific (1 mentions) Taqman gene expression master mix, by Thermo Fisher Scientific (1 mentions) Trap 6, by AnaSpec (1 mentions) Pac 1, by BD (1 mentions) Can get signal solution, by Toyobo (1 mentions) Sds page, by Fujifilm (1 mentions) Chemi lumi one ultra, by Nacalai Tesque (1 mentions)

Spelling variants of this product

GAPDH (5572 citations) Anti-GAPDH (1707 citations) Anti-GAPDH antibody (288 citations) GAPDH antibody (233 citations) GAPDH (14C10) (91 citations) Anti-GAPDH (14C10) (53 citations) GAPDH antibody (14C10) (10 citations) Anti-GAPDH rabbit monoclonal antibody (14C10, (5 citations)

5 protocols using anti gapdh antibody 14c10

Western Blot Analysis of Cell Cycle Proteins

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Proteins in 2 × SDS loading buffer were separated on a 10% SDS polyacrylamide gel and transferred to Immobilon-P PVDF membranes (Merck) using tank blotting (Bio-Rad), with the exception of histone H3 and GAPDH, which were separated on a 15% SDS polyacrylamide gel and transferred using a semi-dry blotting system (ATTO). After blocking with 5% skim milk (Nacalai Tesque), the membranes were incubated with the following primary antibodies: anti-Mcm2 antibody (D7G11; Cell Signaling Technology), anti-GAPDH antibody (14C10; Cell Signaling Technology), anti-histone H3 antibody (1G1, (41 (link))), anti-Cdc6 antibody (sc-9964; Santa Cruz Biotechnology), anti-Cdt1 (D10F11; Cell Signaling Technology), anti-Orc2 antibody (3G6; Santa Cruz Biotechnology), and anti-Cdc45 antibody (D7G6; Cell Signaling Technology). After incubation with peroxidase-conjugated secondary antibodies (GE Healthcare), the signals were detected using ImmunoStar LD (Fujifilm).

Hayashi-Takanaka Y., Hayashi Y., Hirano Y., Miyawaki-Kuwakado A., Ohkawa Y., Obuse C., Kimura H., Haraguchi T, & Hiraoka Y. (2021). Chromatin loading of MCM hexamers is associated with di-/tri-methylation of histone H4K20 toward S phase entry. Nucleic Acids Research, 49(21), 12152-12166.

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Purification and Characterization of Ubiquitin-Conjugating Enzymes

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Human WWP1 cDNA (aa2-922) was purchased from Genscript. The ubiquitin variant gene DNA was synthesized by IDT. The WWP1 and ubiquitin variant gene cDNAs were subcloned into a pGEX6p-2 vector for E. coli expression. The WWP1 K740N and N745S mutations were prepared by QuikChange mutagenesis and the sequence of the entire open reading frames were confirmed by DNA sequencing. Wild-type ubiquitin, ubiquitin-activating enzyme UBA1 (E1), and conjugating enzyme UbcH5b (E2) were prepared as previously described^{25 (link)}. The Colloidal Blue staining kit was purchased from Thermo Fisher. The anti-PTEN antibody (N-19 and A2B1) was purchased from Santa Cruz Biotechnology. The anti-GAPDH antibody (14C10) was purchased from Cell signaling Technology. The transfection reagent Lipofectamine 3000 was purchased from Thermo Fisher Scientific. All other reagents were of the highest quality and obtained commercially from either MilliporeSigma or Thermo Fisher Scientific.

Jiang H., Dempsey D.R, & Cole P.A. (2021). Ubiquitin Ligase Activities of WWP1 Germline Variants K740N and N745S. Biochemistry, 60(5), 357-364.

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IL-1β Measurement in Human Cells

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Human IL-1β enzyme-linked immunosorbent assay kit was from Aviva System Biology. Anti–IL-1β antibody (3A6) and anti-GAPDH antibody (14C10) were from Cell Signaling. Primers for IL-1β (HS01555410), Caspase-1 (HS00354836), TLR2 (HS02621280), TLR4 (HS00152939), MYD88 (HS01573837), and GAPDH (HS02786624) as well as TaqMan Gene Expression Master Mix and complementary DNA Reverse Transcription Kit were from Applied Biosystems. Custom-made pre–IL-1β primer was from Eurofins. FAM-FLICA was from Immunochem Technologies. PAC-1, anti–P-selectin (clone AK4), and anti–IL-1β (clone AS10) were from BD Bioscience. TRAP-6 was from Anaspec. CRP-XL was from Cambcol. All other reagents were from Sigma Aldrich.

Berger M., Maqua H., Lysaja K., Mause S.F., Hindle M.S., Naseem K., Dahl E., Speer T., Marx N, & Schütt K. (2023). Platelets from patients with chronic inflammation have a phenotype of chronic IL-1β release. Research and Practice in Thrombosis and Haemostasis, 8(1), 102261.

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Western Blot Analysis of Signaling Proteins

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Harvested cells were lysed in loading buffer (50 mM Tris‐HCl, pH 6.8, 2% sodium dodecylsulfate (SDS), 10% glycerol, 6% 2‐mercaptoethanol, 0.0025% bromophenol blue), subjected to SDS‐PAGE (Wako), and then transferred to a PVDF membrane. The following primary antibodies were diluted in Can Get Signal Solution (TOYOBO): anti‐LMP1 antibody (S12, 1:50), anti‐GAPDH antibody (14C10; Cell Signaling Technology [CST], 1:2000), anti‐TBK1 antibody (CST), anti‐phospho‐TBK1/NAK (Ser172) antibody (D52C2, 1:1000; CST), and anti‐STING antibody (D2P2F, 1:1000; CST). As the secondary antibody, HRP‐conjugated goat anti‐mouse IgG (BioSource) or HRP‐linked anti‐rabbit IgG (CST) was used at a dilution of 2000‐5000. Forte (Merck) or Chemi‐Lumi One Ultra (Nacalai Tesque) was used as the HRP substrate, and EZ‐Analyzer (ATTO) was used to detect luminescence, which was also photographed.

Miyagi S., Watanabe T., Hara Y., Arata M., Uddin M.K., Mantoku K., Sago K., Yanagi Y., Suzuki T., Masud H.M., Kawada J., Nakamura S., Miyake Y., Sato Y., Murata T, & Kimura H. (2021). A STING inhibitor suppresses EBV‐induced B cell transformation and lymphomagenesis. Cancer Science, 112(12), 5088-5099.

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Western Blot Analysis of Cell Cycle Proteins

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Proteins in 2× SDS loading buffer were separated on a 10% SDS polyacrylamide gel and transferred to Immobilon-P PVDF membranes (Merck) using tank blotting (Bio-Rad), with the exception of histone H3 and GAPDH, which were separated on a 15% SDS polyacrylamide gel and transferred using a semi-dry blotting system (ATTO). After blocking with 5% skim milk (Nacalai Tesque), the membranes were incubated with the following primary antibodies: anti-Mcm2 antibody (D7G11; Cell Signaling Technology), anti-GAPDH antibody (14C10; Cell Signaling Technology), anti-histone H3 antibody (1G1, ( 41)), anti-Cdc6 antibody (sc-9964; Santa Cruz Biotechnology), anti-Cdt1 (D10F11; Cell Signaling Technology), anti-Orc2 antibody (3G6; Santa Cruz Biotechnology), and anti-Cdc45 antibody (D7G6; Cell Signaling Technology). After incubation with peroxidase-conjugated secondary antibodies (GE Healthcare), the signals were detected using ImmunoStar LD (Fujifilm).

Hayashi‐Takanaka Y., Hayashi Y., Hirano Y., Miyawaki‐Kuwakado A., Ohkawa Y., Obuse C., Kimurâ H., Haraguchi T., & Hiraoka Y. (2021). Chromatin loading of MCM hexamers is associated with di-/tri-methylation of histone H4K20 toward S phase entry.

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