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Product Name GAPDH (Loading Control) Chinese Name 3-磷酸甘油醛脱氢酶(内参)抗体 Alias 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 dehydrogenase muscle; KNC-NDS6; MGC102544; MGC102546; MGC103190; MGC103191; MGC105239; MGC127711; MGC88685; OCAS, p38 component; OCT1 coactivator in S phase, 38-KD component; wu:fb33a10. literatures Specific References (53) | SL10900R has been referenced in 53 publications.Product Type Internal reference anti Research Area Tumour Cell biology immunology Signal transduction The new supersedes the old Immunogen Species Rabbit Clonality Polyclonal React Species Human, Mouse, Rat, Applications WB=1:10000-200000 IHC-P=1:100-500 IHC-F=1:100-500 ICC=1:100 IF=1:100-500 (Paraffin sections need antigen repair)
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.Theoretical molecular weight 38kDa Cellular localization The nucleus cytoplasmic The cell membrane Form Liquid Concentration 1mg/ml immunogen Recombinant human GAPDH full length protein Lsotype IgG Purification affinity purified by Protein A Buffer Solution 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Storage Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. Attention This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. PubMed PubMed Product Detail 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:
P04406
Gene ID:
2597
Database links:Entrez Gene: 374193 Chicken
Entrez Gene: 2597 Human
Entrez Gene: 100042025 Mouse
Entrez Gene: 14433 Mouse
Entrez Gene: 317743 Zebrafish
Omim: 138400 Human
SwissProt: P00356 Chicken
SwissProt: P04406 Human
SwissProt: P16858 Mouse
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Internal reference anti。
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.Product Picture Sample: 293T(human) cell lysate at 30ug;
Primary:
Lane1: Anti-GAPDH (SL10900R) at 1/2000 dilution
Lane2: Anti-GAPDH (SL10900R) at 1/10000 dilution
Lane3: Anti-GAPDH (SL10900R) at 1/40000 dilution
Lane4: Anti-GAPDH (SL10900R) at 1/80000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 38 kD
Observed band size: 38kD
Sample:
Lane 1: Human HeLa cell lysates
Lane 2: Human U-2 OS cell lysates
Lane 3: Mouse NIH/3T3 cell lysates
Lane 4: Rat RSC96 cell lysates
Primary: Anti-GAPDH (SL10900R) at 1/20000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 37 kDa
Observed band size: 37 kDa
Sample: 293T (Human) Lysate at 30 ug
A549(Human) Lysate at 30 ug
Primary: Anti-GAPDH (SL10900R-HRP) at 1/1000 dilution
Predicted band size: 38 kD
Observed band size: 38 kD
Sample:
A549 Cell (Human) Lysate at 40 ug
A431 Cell (Human) Lysate at 40 ug
NIH/3T3 Cell (Mouse) Lysate at 40 ug
Primary: Anti-GAPDH (SL10900R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 38 kD
Observed band size: 36 kD
Sample:
Heart (Mouse) Lysate at 40 ug
Cerebrum (Mouse) Lysate at 40 ug
Liver (Mouse) Lysate at 40 ug
Kidney (Mouse) Lysate at 40 ug
Primary: Anti- GAPDH (SL10900R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 38 kD
Observed band size: 38 kD
Sample:
Lane 1: SiHa (Human) Cell Lysate at 30 ug
Lane 2: NIH/3T3(Mouse) Cell Lysate at 30 ug
Lane 3: Large intestine (Mouse) Lysate at 40 ug
Lane 4: Cerebrum (Rat) Lysate at 40 ug
Lane 5: Cerebrum (Mouse) Lysate at 40 ug
Lane 6: Testis (Rat) Lysate at 40 ug
Lane 7: Testis (Mouse) Lysate at 40 ug
Lane 8: Kidney (Mouse) Lysate at 40 ug
Lane 9: HUVEC (Human) Cell Lysate at 30 ug
Lane 10: A549 (Human) Cell Lysate at 30 ug
Lane 11: MCF-7 (Human) Cell Lysate at 30 ug
Primary: Anti-GAPDH (SL10900R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 36 kD
Observed band size: 36 kD
Sample:
293T (Human) Lysate at 40 ug
Primary:
Anti-GAPDH (SL10900R) at 1/2000~1/20000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 38 kD
Observed band size: 36 kD
Sample:
293T(Human) Cell Lysate at 30 ug
Primary: Anti-GAPDH (SL10900R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 38 kD
Observed band size: 38 kD
Sample:
H9C2(Rat) Cell Lysate at 30 ug
Primary: Anti-GAPDH (SL10900R) at 1/2000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 38 kD
Observed band size: 38 kD
Paraformaldehyde-fixed, paraffin embedded (rat colon); 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 (Loading Control)) Polyclonal Antibody, Unconjugated (SL10900R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (mouse colon); 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 (Loading Control)) Polyclonal Antibody, Unconjugated (SL10900R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (Human kidney); 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) Polyclonal Antibody, Unconjugated (SL10900R) at 1:2000 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.Paraformaldehyde-fixed, paraffin embedded (human 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 (Loading Control)) Polyclonal Antibody, Unconjugated (SL10900R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (human colon); 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 (Loading Control)) Polyclonal Antibody, Unconjugated (SL10900R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (human lung carcinoma); 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 (Loading Control)) Polyclonal Antibody, Unconjugated (SL10900R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.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 (Loading Control)) Polyclonal Antibody, Unconjugated (SL10900R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Bought notes(bought amounts latest0)
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