Mouse Anti-GAPDH (Loading Control) antibody

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38 kDa BFA-dependent ADP-ribosylation substrate; Aging-associated gene 9 protein; BARS-38; cb609; EC 1.2.1.12; G3PD; G3PDH; GAPD; Glyceraldehyde 3 phosphate dehydrogenase;Glyceraldehyde 3 phosphate dehydrogenase liver;Glyceraldehyde 3 phosphate dehydrogen

Cat:

SLM0978M

Species Reactivity：

Human,Mouse,Rat,(predicted: Rabbit,)

Immunogen：

Full length GAPDH protein purified from rabbit muscle

Format：

Liquid

Storage instructions：

Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Concentration：

1mg/ml

Clonality：

Monoclonal

Isotype：

IgG

Applications：

WB=1:1000-5000IHC-P=1:100-500IHC-F=1:100-500ICC=1:100IF=1:100-500（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.

Host：

Mouse

Product Overview：

Sample: Placenta (Mouse) Lysate at 40 ug293T Cell Lysate at 40 ugPrimary: Anti-GAPDH(SLM0978M)at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilutionPredicted band size: 38kDObserved band size: 38kDSample: Bladder(Rat)Lysate at 40 ugPrimary: Anti-GAPDH(SLM0978M)at 1/300 dilutionSecondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilutionPredicted band size: 38kDObserved band size: 38kDSample: A549(Human) Cell Lysate at 30 ug293T(Human) Cell Lysate at 30 ugHepG2(Human) Cell Lysate at 30 ugHela(Human) Cell Lysate at 30 ugPrimary: Anti- GAPDH(3E12) (SLM0978M) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilutionPredicted band size: 38 kDObserved band size: 38 kDSample:Liver(Mouse)Lysate at 40 ugPrimary: Anti-GAPDH(SLM0978M)at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilutionPredicted band size: 38kDObserved band size: 38kDSample: Lane 1: Hela (Human) Cell Lysate at 30 ugLane 2: Huvec (Human) Cell Lysate at 30 ugLane 3: A549 (Human) Cell Lysate at 30 ugLane 4: MCF-7 (Human) Cell Lysate at 30 ugPrimary: Anti-GAPDH (SLM0978M) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilutionPredicted band size: 36 kDObserved band size: 36 kDSample: Cerebrum (Mouse) Lysate at 40 ugPrimary: Anti-GAPDH(3E12)-Loading Control (SLM0978M) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilutionPredicted band size: 38 kDObserved band size: 38 kDParaformaldehyde-fixed, paraffin embedded (Rat pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH(3E12)-Loading Control) Polyclonal Antibody, Unconjugated (SLM0978M) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining. Tissue/cell: HUVEC cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, SLC0005) at 37°C for 20 min; Antibody incubation with (GAPDH(3E12)-Loading Control) Polyclonal Antibody, Unconjugated (SLM0978M) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody (SL0296G-FITC) at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

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Unit:

Price: $136.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
Feedback Wall

Loading Control
Glyceraldehyde 3 phosphate dehydrogenase (GAPDH) is well known as one of the key enzymes involved in glycolysis. As well as functioning as a glycolytic enzyme in cytoplasm, recent evidence suggests that mammalian GAPDH is also involved in a great number of intracellular proceses such as membrane fusion, microtubule bundling, phosphotransferase activity, nuclear RNA export, DNA replication, and DNA repair. During the last decade a lot of data appeared concerning the role of GAPDH in different pathologies including prostate cancer progression, programmed neuronal cell death, age related neuronal diseases, such as Alzheimer's and Huntington's disease. GAPDH is expressed in all cells. It is constitutively expressed in almost all tissues at high levels. There are however some physiological factors such as hypoxia and diabetes that increase GAPDH expression in certain cell types. GAPDH molecule is composed of four 36kDa subunits.

Function:
Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC. Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate.

Subunit:
Homotetramer. Interacts with TPPP; the interaction is direct. Interacts (when S-nitrosylated) with SIAH1; leading to nuclear translocation. Interacts with RILPL1/GOSPEL, leading to prevent the interaction between GAPDH and SIAH1 and prevent nuclear translocation. Interacts with EIF1AD, USP25, PRKCI and WARS.

Subcellular Location:
Cytoplasm, cytosol. Nucleus. Cytoplasm, perinuclear region. Membrane. Note=Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal. Postnuclear and Perinuclear regions.

Post-translational modifications:
S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
ISGylated (Probable).
Sulfhydration at Cys-152 increases catalytic activity.

Similarity:
Belongs to the glyceraldehyde-3-phosphate dehydrogenase family.

SWISS:
P41286

Gene ID:
100009074

Database links:

Entrez Gene: 374193 Chicken

Entrez Gene: 2597 Human

Entrez Gene: 100042025 Mouse

Entrez Gene: 14433 Mouse

Entrez Gene: 24383 Rat

Entrez Gene: 685186 Rat

Entrez Gene: 317743 Zebrafish

Omim: 131680 Human

SwissProt: P00356 Chicken

SwissProt: P0886 Human

SwissProt: P16858 Mouse

SwissProt: P04797 Rat

SwissProt: Q5XJ10 Zebrafish

GAPDH蛋白几乎在所有组织中都高水平表达，广泛用作Western blot蛋白质标准化的内参,是很好的内参抗体。
GAPDH 作为管家基因在同种细胞或者组织中的蛋白质表达量一般是恒定的。在实验中，可能存在总蛋白浓度测定不准确；或者蛋白质样品在电泳前上样时产生的样品间的操作误差；这些误差需要通过测定每个样品中实际转到膜上的GAPDH的含量来进行校正，所以一般的western实验都需要进行内参设置。具体校正的方法就是将每个样品测得的目的蛋白含量与本样品的GAPDH含量相除，得到每个样品目的蛋白的相对含量。然后才进行样品与样品之间的比较。
甘油醛-3-磷酸脱氢酶（Glyceraldehyde 3 phosphate dehydrogenase，GAPDH）是糖酵解（glycolysis）过程中的关键酶。除了在胞质中作为糖酵解的酶以外，有证据表明哺乳动物细胞中的GAPDH参与了多种胞内生化过程，包括膜融合（membrane fusion）、微管成束（microtubule bundling）、磷酸转移酶（phosphotransferase）激活、核内RNA出核、DNA复制与DNA修复。一些生理因素，诸如低氧（hypoxia）和尿糖（diabetes），可以增加GAPDH在特定细胞中的表达。GAPDH存在于几乎所有的组织中，以高水平持续表达。
GAPDH（甘油醛-3-磷酸脱氢酶）是参与糖酵解的一种关键酶，由4个30－40kDa的亚基组成. Picture

Sample: Bladder(Rat)Lysate at 40 ug
Primary: Anti-GAPDH(SLM0978M)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilution
Predicted band size: 38kD
Observed band size: 38kD

Sample:
A549(Human) Cell Lysate at 30 ug
293T(Human) Cell Lysate at 30 ug
HepG2(Human) Cell Lysate at 30 ug
Hela(Human) Cell Lysate at 30 ug
Primary: Anti- GAPDH(3E12) (SLM0978M) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution
Predicted band size: 38 kD
Observed band size: 38 kD

Sample:Liver(Mouse)Lysate at 40 ug
Primary: Anti-GAPDH(SLM0978M)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution
Predicted band size: 38kD
Observed band size: 38kD

Sample:
Lane 1: Hela (Human) Cell Lysate at 30 ug
Lane 2: Huvec (Human) Cell Lysate at 30 ug
Lane 3: A549 (Human) Cell Lysate at 30 ug
Lane 4: MCF-7 (Human) Cell Lysate at 30 ug
Primary: Anti-GAPDH (SLM0978M) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution
Predicted band size: 36 kD
Observed band size: 36 kD

Sample:
Cerebrum (Mouse) Lysate at 40 ug
Primary: Anti-GAPDH(3E12)-Loading Control (SLM0978M) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution
Predicted band size: 38 kD
Observed band size: 38 kD

Paraformaldehyde-fixed, paraffin embedded (Rat pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH(3E12)-Loading Control) Polyclonal Antibody, Unconjugated (SLM0978M) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.

Tissue/cell: HUVEC cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, SLC0005) at 37°C for 20 min; Antibody incubation with (GAPDH(3E12)-Loading Control) Polyclonal Antibody, Unconjugated (SLM0978M) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody (SL0296G-FITC) at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Product Feedback Wall

Specific References (74) | SLM0978M has been referenced in 74 publications.

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[IF=5.88] Shuai Chen. et al. Tenogenic adipose-derived stem cell sheets with nanoyarn scaffolds for tendon regeneration. Mat Sci Eng SLCMater. 2021 Feb;119:111506 WB ; Rabbit.

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[IF=4.534] Ji-Lun Cheng. et al. LncRNA KCNQ1OT1 regulates the invasion and migration of hepatocellular carcinoma by acting on S1PR1 through miR-149. Cancer Gene Ther. 2020 Aug;:1-14 WB ; Human.

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[IF=4.872] Shaofeng Wu. et al. Protective effects of curcumin on ATO-induced nephrotoxicity in ducks in relation to suppressed autophagy, apoptosis and dyslipidemia by regulating oxidative stress. Ecotox Environ Safe. 2021 Aug;219:112350 WB ; Duck.

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[IF=3.06] Wang, Huihui. et al. Anti-inflammatory and Antioxidant Effects of Pyrroloquinoline Quinone in L-NAME-Induced Preeclampsia-Like Rat Model. 2021 Sep 20 WB ; Rat.

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[IF=6.244] Pei Dai. et al. Gimap5 Inhibits Lung Cancer Growth by Interacting With M6PR. Front Oncol. 2021; 11: 699847 WB ; Human.

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[IF=5.187] Mingzhu Jiang. et al. Blocking AURKA with MK-5108 attenuates renal fibrosis in chronic kidney disease. Bba-Mol Basis Dis. 2021 Nov;1867:166227 WB ; Rat.

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[IF=4.486] Zhuohong Li. et al. Anlotinib exerts anti-cancer efficiency on lung cancer stem cells in vitro and in vivo through reducing NF-κB activity. 2021 May 06 WB ; Mouse.

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[IF=3.056] Shaofeng Wu. et al. Evaluation of toxic effects induced by arsenic trioxide or/and antimony on autophagy and apoptosis in testis of adult mice. 2021 May 20 WB ; Mouse.

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[IF=4.225] Hao Ding. et al. Effects of Real-Ambient PM2.5 Exposure on Lung Damage Modulated by Nrf2−/−. Front Pharmacol. 2021; 12: 662664 WB ; Mouse.

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[IF=2.363] Wang Li. et al. Trilobatin Induces Apoptosis and Attenuates Stemness Phenotype of Acquired Gefitinib Resistant Lung Cancer Cells via Suppression of NF-κB Pathway. 2021 Apr 16 WB ; Human.

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[IF=3.876] Daisuke Fujioka. et al. Group V Secretory Phospholipase A2 Regulates Endocytosis of Acetylated LDL by Transcriptional Activation of PGK1 in RAW264.7 Macrophage Cell Line. 2021 Mar 27 WB ; Mouse.

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[IF=5.606] Luo Ping. et al. IL-37 inhibits M1-like macrophage activation to ameliorate temporomandibular joint inflammation through the NLRP3 pathway. Rheumatology. 2020 Oct;59(10):3070-3080 WB ; Human.

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[IF=5.201] Guo Qiang. et al. PAQR3 Inhibits Non-small Cell Lung Cancer Growth by Regulating the NF-κB/p53/Bax Axis. Front Cell Dev Biol. 2020 Oct;8:1060 WB ; Human.

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[IF=3.267] Seona Cho. et al. Viburnum stellato-tomentosum Extract Suppresses Obesity and Hyperglycemia through Regulation of Lipid Metabolism in High-Fat Diet-Fed Mice. Molecules. 2021 Jan;26(4):1052 WB ; Mouse.

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[IF=3.212] Gaolong Zhong. et al. The protective role of autophagy against arsenic trioxide-induced cytotoxicity and ROS-dependent pyroptosis in NCTSLC1469 cells. J Inorg Biochem. 2021 Apr;217:111396 WB ; Mouse.

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[IF=6.551] Yukun Fang. et al. Activation of the ROS/HO-1/NQO1 signaling pathway contributes to the copper-induced oxidative stress and autophagy in duck renal tubular epithelial cells. Sci Total Environ. 2021 Feb;757:143753 WB ; Duck.

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[IF=1.931] Yongtao Li. et al. Effects of epigallocatechin gallate (EGCG) on the biological properties of human dental pulp stem cells and inflammatory pulp tissue. Arch Oral Biol. 2021 Mar;123:105034 WB ; Human.

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[IF=6.551] Gaolong Zhong. et al. Arsenic or/and antimony induced mitophagy and apoptosis associated with metabolic abnormalities and oxidative stress in the liver of mice. Sci Total Environ. 2021 Jul;777:19282 WB ; Mouse.

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[IF=3.998] Xiao-Yi Zhang. et al. Dietary capsaicin normalizes CGRP peptidergic DRG neurons in experimental diabetic peripheral neuropathy. Sci Rep-Uk. 2021 Jan;11(1):1-19 WB ; Rat.

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[IF=3.297] Hongyi Qu. et al. The effects of vasoactive intestinal peptide on RANKL-induced osteoclast formation. Ann Transl Med. 2021 Jan; 9(2): 127 WB ; Rat.

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[IF=2.842] Chen T et al. p53 Mediates GnRH Secretion via Lin28/let-7 System in GT1-7 CellsDiabetes Metab Syndr Obes.2020 Dec 1;13:4681-4688. WB ; Mouse.

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[IF=3.647] Weiyi Zhanget al. Cinnamaldehyde changes the dynamic balance of glucose metabolism by targeting ENO1. Life Sci . 2020 Oct 1;258:118151. other ;

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[IF=3.337] Xuesong Wanget al. LncRNA NEAT1 Regulates 5-Fu Sensitivity, Apoptosis and Invasion in Colorectal Cancer Through the MiR-150-5p/CPSF4 Axis. Onco Targets Ther . 2020 Jul 1;13:6373-6383. WB ; Human.

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[IF=2.13] Wei, Ting, et al. "The role of N-acetylglucosaminyltransferases V in the malignancy of human hepatocellular carcinoma." Experimental and molecular pathology (2012). WB ; Human.

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