Gapdh 14c10 rabbit mab

Manufactured by Cell Signaling Technology

Sourced in United States

GAPDH (14C10) Rabbit mAb is a laboratory reagent used to detect the presence and quantify the levels of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein in biological samples. GAPDH is a ubiquitous enzyme involved in the glycolytic pathway and is commonly used as a loading control or reference protein in various biochemical and cell biology experiments.

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Lab products found in correlation

M per mammalian protein extraction reagent, by Thermo Fisher Scientific (1 mentions) Micro bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Phosphatase inhibitor cocktail b, by Santa Cruz Biotechnology (1 mentions) Phosphatase inhibitor cocktail a, by Santa Cruz Biotechnology (1 mentions) Supersignal west femto maximum sensitivity substrate, by Thermo Fisher Scientific (1 mentions) Image studio lite 5, by LI COR (1 mentions) Tissuelyser lt, by Qiagen (1 mentions) Proteinase and phosphatase inhibitors, by Merck Group (1 mentions) Bca assay, by Thermo Fisher Scientific (1 mentions) Goat anti rabbit igg hrp, by Santa Cruz Biotechnology (1 mentions) Odyssey fc imaging system, by LI COR (1 mentions) N6 2 o dibutyryladenosine 3 5 cyclic monophosphate bt2camp, by Merck Group (1 mentions) 22 r hydroxycholesterol, by Merck Group (1 mentions) 17β estradiol, by Merck Group (1 mentions) Progesterone, by Merck Group (1 mentions) Fatty acid free bovine serum albumin, by Merck Group (1 mentions) Phospho p38 mapk thr180 tyr182 antibody, by Cell Signaling Technology (1 mentions) Ipvh00010, by Cytiva (1 mentions) Protease inhibitor, by Merck Group (1 mentions) Nitrocellulose membrane, by Cytiva (1 mentions)

Spelling variants of this product

GAPDH (5572 citations) GAPDH (14C10) (91 citations) GAPDH rabbit mAb (19 citations) Anti–GAPDH (14C10) Rabbit mAb (9 citations) GAPDH mAb (6 citations) Rabbit mABs (2 citations)

8 protocols using gapdh 14c10 rabbit mab

Biochemical Analysis of Phosphatase Activity

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4G10 Platinum and Anti-Phosphotyrosine Antibody (05-1050) were obtained from EDM Millipore. Rabbit-β-Tubulin antibody (H-235), Phosphatase Inhibitor Cocktail A, and Phosphatase Inhibitor Cocktail B were obtained from Santa Cruz Biotechnologies. 6, 8-Difluoro-4-Methylumbelliferyl Phosphate (DiFMUP), Alexa Fluor 555 goat anti-rabbit IgG (A-21428), and Alexa Fluor 647 goat anti-mouse IgG (A-21236) were obtained from Thermo Fisher Scientific (Invitrogen). GAPDH (14C10) Rabbit mAb was obtained from Cell Signaling Technology. Mammalian Protein Extraction Reagent (M-PER) and Micro BCA Protein Assay Kit were obtained from Thermo Fisher Scientific. Recombinant human PTP1B and PP1 were obtained from Novus Biologicals.

Cornejo N.R., Amofah B., Lipinski A., Langlais P.R., Ghosh I, & Jewett J.C. (2022). Direct intracellular delivery of benzene diazonium ions as observed by increased tyrosine phosphorylation. Biochemistry, 61(8), 656-664.

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Murine Heart Tissue Protein Isolation

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Protein isolation of murine heart tissue was performed with the TissueLyser LT (Qiagen) using RIPA buffer with proteinase and phosphatase inhibitors (Sigma-Aldrich). Samples were incubated at 4°C for 30 min followed by sonification. Protein concentrations were measured via BCA assay (Thermo Fisher Scientific) according to manufacturer’s protocol.
100 µg total protein was separated on a 12% SDS-gel and transferred on nitrocellulose membrane in transfer buffer containing 18% methanol for 1 h. Membranes were blocked in 5% milk in TBS containing 0.05% Tween-20 for 1 h, afterwards primary antibodies were incubated over-night at 4°C (FGF23-6310 Goat mAb, dilution 1:500, Immutopics; GAPDH 14C10 Rabbit mAb, dilution 1:1,000, Cell Signaling Technology). After washing, secondary antibodies were incubated for 1 h at room temperature (Goat anti-rabbit IgG HRP, dilution 1:1,000, Santa Cruz; Donkey anti-goat IgG HRP, dilution 1:2000, R and D Systems). ECL development was done with the SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Fisher Scientific) and signals were detected with the Odyssey FC Imaging System (LI-COR Bioscience). Expression levels were quantified with the Image Studio Lite 5.2 software (LI-COR Bioscience).

Eitner F., Richter B., Schwänen S., Szaroszyk M., Vogt I., Grund A., Thum T., Heineke J., Haffner D, & Leifheit-Nestler M. (2022). Comprehensive Expression Analysis of Cardiac Fibroblast Growth Factor 23 in Health and Pressure-induced Cardiac Hypertrophy. Frontiers in Cell and Developmental Biology, 9, 791479.

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Steroid Hormone Signaling Pathway

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22(R)-hydroxycholesterol, 17β-estradiol, N6,2’-O-dibutyryladenosine 3’,5’-cyclic monophosphate (Bt₂cAMP), progesterone, 20α-OHP, 22(R)-hydroxycholesterol and fatty acid-free bovine serum albumin were supplied by Sigma-Aldrich (St. Louis, MO). 20α-Hydroxyprogesterone [1,2-³H(N)] (40–60 Ci/mmol; 1.48–2.22 TBq/mmol) and progesterone [1,2-³H] (40–60 Ci/mmol; 1.48–2.22 TBq/mmol) were purchased from American Radiolabeled Chemicals, Inc. (St. Louis, MO). Pregnenolone, [7-³H(N)] (10–25 Ci/mmol; 370–925 GBq/mmol) was supplied by PerkinElmer (Waltham, MA). StAR (D10H12) XP^® rabbit mAb, GAPDH (14C10) rabbit mAb, and MeCP2 (D4F3) XP™ rabbit mAb were obtained from Cell Signaling Technology, Inc. (Danvers, MA). Purified anti-SF-1 polyclonal antibody (07–618) was purchased from EMD Millipore (Billerica, MA). All other chemicals used were of analytical grade.

Hu Z., Shen W.J., Kraemer F.B, & Azhar S. (2017). Regulation of adrenal and ovarian steroidogenesis by miR-132. Journal of molecular endocrinology, 59(3), 269-283.

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Western Blot Analysis of Signaling Pathways

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One million U937 cells were stimulated, washed in PBS, and lysed in buffer (4% SDS, 40 mM HEPES [pH 7.4, 10 mM DTT] supplemented with protease inhibitors [Sigma‐Aldrich, 4693159001]). Samples were centrifuged (16,000 g, 10 min), Li‐LDS sample buffer was added to a final concentration of 1×, and the supernatant was incubated (5 min, 95°C). Proteins were separated on 12% Novex Tris‐glycine gels (Thermo Fisher Scientific, XP00120BOX) and transferred onto PVDF membranes (Merck Millipore, IPVH00010) or Nitrocellulose membranes (Amersham, 10600002). Membranes were blocked in 5% BSA in PBST, and antibodies were diluted in 2% BSA in PBST. Antibodies used for immunoblotting were as follows (diluted 1:1,000): phospho‐p38 MAPK (Thr180/Tyr182) antibody (Cell Signaling, 9211), GAPDH (14C10) rabbit mAb (Cell Signalling, 2118), p38 MAPK (R&D, AF8691), ARHGEF18 (Sigma, HPA042689), MAP3K7 (R&D, MAB5307), FOSB (R&D, AF2214) and anti‐rabbit IgG, HRP‐linked antibody (Cell Signaling, 7074).

Frauenstein A., Ebner S., Hansen F.M., Sinha A., Phulphagar K., Swatek K., Hornburg D., Mann M, & Meissner F. (2021). Identification of covalent modifications regulating immune signaling complex composition and phenotype. Molecular Systems Biology, 17(7), e10125.

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Antibody Panel for Stem Cell Analysis

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The following antibodies were used for Western blot and immunocytochemistry: Anti-Oct4 (Abcam, ab19857), anti-SOX17 antibody [3B10] (Abcam, ab 60721), anti-FOXA2 antibody (Abcam, ab 84990), anti-HNF-4-alpha antibody [K9218] (Abcam, ab 41898), anti-PARP antibody (Cell Signaling Technology, 9542), anti-caspase-3 antibody (Cell Signaling Technology, 9662), Nanog (1E6C4) Mouse mAb (Cell Signaling Technology, 4893), Sox2 (L73B4) Mouse mAb (Cell Signaling Technology, 4195), GAPDH (14C10) Rabbit mAb (Cell Signaling Technology, 2118), and anti-PAX-6 (Biolegend, 901301).

Varghese D.S., Parween S., Ardah M.T., Emerald B.S, & Ansari S.A. (2017). Effects of Aminoglycoside Antibiotics on Human Embryonic Stem Cell Viability during Differentiation In Vitro. Stem Cells International, 2017, 2451927.

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D. latiflorus Leaf Extract Effects

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D. latiflorus leaves were purchased from Maoshennongye Co., Ltd. (Shandong, China). HepG2 cell line was obtained from the National Infrastructure of Cell Line Resource, Peking Union Medical College (Beijing, China). Fetal bovine serum (FBS) Uruguay was purchased from Biowest (Nuaillé, France). Dulbecco's modified Eagle's medium (DMEM) with high glucose, liquid and other cell culture reagents were purchased from HyClone (Logan, UT, USA). AKT antibody, phospho-AKT (Ser473) (D9E) XP rabbit mAb, FOXO1 (C29H4) rabbit mAb, phospho-FOXO1 (Ser256) antibody, GSK-3β (D5C5Z) XP rabbit mAb, phospho-GSK-3β (Ser9) (D85E12) XP rabbit mAb, and GAPDH (14C10) rabbit mAb were purchased from Cell Signaling Technology (Beverly, MA, USA). The insulin growth factor 1 receptor (IGF1R) inhibitor GSK1904529A was purchased from Selleck Chemicals (Houston, TX, USA).

Luo K., Huang W., Qiao L., Zhang X., Yan D., Ning Z., Ma C., Dang H., Wang D., Guo H., Xie L, & Cheng J. (2021). Dendrocalamus latiflorus and its component rutin exhibit glucose-lowering activities by inhibiting hepatic glucose production via AKT activation. Acta Pharmaceutica Sinica. B, 12(5), 2239-2251.

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RANKL-Induced RAW264.7 Cells Proteome Analysis

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RANKL‐stimulated RAW264.7 cells were lysed in CytoBuster^TM Protein Extraction Reagent (Millipore, Cat# 71009) supplemented with protease and phosphatase inhibitors. The lysates were denatured, separated in a Bolt™ 4–12% Bis‐Tris gel (Invitrogen, Cat# NW04120BOX) and transferred to a membrane using an iBlot™ 2 PVDF Gel Transfer Stack (Invitrogen, Cat# IB24001). The membrane was blocked with 5% Blotting Grade Blocker Non Fat Dry Milk (BioRad, Cat# 1706404XTU) and incubated overnight with Human/Mouse/Rat Galectin‑3 Antibody (R&D Systems, Cat# AF1197), Anti‐LAMP‐1 (CD107a) Antibody (Sigma Aldrich, Cat# AB2971) and GAPDH (14C10) Rabbit mAb (Cell Signaling Technology, Cat# 2118) in blocking buffer. The cells were, then, incubated with HRP‐linked secondary antibodies, and the protein bands were detected using the chemiluminescent HRP substrate SuperSignal™ West Pico PLUS (Thermo Scientific, Cat# 34579).

Duarte C., Yamada C., Garcia C., Akkaoui J., Ho A., Nichols F, & Movila A. (2022). Crosstalk between dihydroceramides produced by Porphyromonas gingivalis and host lysosomal cathepsin B in the promotion of osteoclastogenesis. Journal of Cellular and Molecular Medicine, 26(10), 2841-2851.

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Western Blot Analysis of Key Signaling Proteins

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Cell pellets were resuspended in lysis buffer RIPA, and protein concentration was assessed by the Qubit Protein Assay Kit. Protein lysates were separated on SDS-PAGE gels and blotted onto polyvinylidene difluoride (PVDF) membranes (Millipore). PVDF membranes were incubated with the relevant primary antibody and corresponding secondary antibody. Images were processed and acquired using chemiluminescence (Pierce ECL). Antibodies used in this study were: phospho-Stat5 (Tyr694) (C11C5) rabbit monoclonal antibody (#9359, Cell Signaling Technology), Stat5 (D2O6Y) rabbit mAb (#94205T, Cell Signaling Technology), phospho-AMPKa (Thr172) (40H9) rabbit monoclonal antibody (#2535, Cell Signaling Technology), AMPKa (D5A2) Rabbit mAb (#5831T, Cell Signaling Technology), phospho-mTOR (Ser2448) antibody (#2971, Cell Signaling Technology), anti-GATM antibody (#HPA026077, Sigma), GAPDH (14C10) Rabbit mAb (#2118S, Cell Signaling Technology), and antirabbit IgG, HRP-linked antibody (Cell Signaling Technology).

Zhang Y., Newsom K.J., Zhang M., Kelley J.S, & Starostik P. (2022). GATM-Mediated Creatine Biosynthesis Enables Maintenance of FLT3-ITD-Mutant Acute Myeloid Leukemia. Molecular cancer research : MCR, 20(2).

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