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3-磷酸甘油醛脫氫酶

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中文名稱 3-磷酸甘油醛脫氫酶
別    名 Glyceraldehyde-3-phosphate dehydrogenase.  

 

研究領(lǐng)域 細(xì)胞生物  免疫學(xué)  
抗體來(lái)源 Rabbit
克隆類(lèi)型 Polyclonal
交叉反應(yīng) Human, Rat,  (predicted: Mouse, )
產(chǎn)品應(yīng)用 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 Flow-Cyt=1ug/Test ICC=1:100 IF=1:100-500 (石蠟切片需做抗原修復(fù))
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.
分 子 量 37kDa
細(xì)胞定位 細(xì)胞核 細(xì)胞漿 細(xì)胞膜 
性    狀 Liquid
濃    度 1mg/ml
免 疫 原 KLH conjugated synthetic peptide derived from the middle of human GAPDH:231-335/335 
亞    型 IgG
純化方法 affinity purified by Protein A
儲(chǔ) 存 液 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
保存條件 Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
PubMed PubMed
產(chǎn)品介紹 Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is well known as one of the key enzymes involved in glycolysis. It catalyzes an important energy-yielding step in carbohydrate metabolism, thereversible oxidative phosphorylation of glyceraldehyde-3-phosphatein the presence of inorganic phosphate and nicotinamide adenine dinucleotide (NAD). Recent evidence suggests that it also is involved in a number of cellular processes such as membrane fusion, phosphotransferase activity, DNA replication and repair, and nuclear RNA export (1). GAPDH has also been implicated in playing a role in different pathologies such as cancer progression, apoptosis, and neuronal diseases such as Alzheimer’s and Huntington’s disease (2). GAPDH is constitutively expressed at high levels in almost all tissues and cell lines making it ideal for use as a loading control marker in immunoblots.

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: 2597 Human

Entrez Gene: 100042025 Mouse

Entrez Gene: 14433 Mouse

Entrez Gene: 24383 Rat

Entrez Gene: 685186 Rat

Omim: 138400 Human

SwissProt: P04406 Human

SwissProt: P16858 Mouse

SwissProt: P04797 Rat

Unigene: 544577 Human

Unigene: 592355 Human

Unigene: 598320 Human

Unigene: 304088 Mouse

Unigene: 129558 Rat

Unigene: 91450 Rat



Important Note:
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.

甘油醛-3-磷酸脫氫酶(Glyceraldehyde 3 phosphate dehydrogenase,GAPDH)是糖酵解(glycolysis)過(guò)程中的關(guān)鍵酶。除了在胞質(zhì)中作為糖酵解的酶以外,有證據(jù)表明哺乳動(dòng)物細(xì)胞中的GAPDH參與了多種胞內(nèi)生化過(guò)程,包括膜融合(membrane fusion)、微管成束(microtubule bundling)、磷酸轉(zhuǎn)移酶(phosphotransferase)激活、核內(nèi)RNA出核、DNA復(fù)制與DNA修復(fù)。一些生理因素,諸如低氧(hypoxia)和尿糖(diabetes),可以增加GAPDH在特定細(xì)胞中的表達(dá)。GAPDH存在于幾乎所有的組織中,以高水平持續(xù)表達(dá)。
GAPDH(甘油醛-3-磷酸脫氫酶)是參與糖酵解的一種關(guān)鍵酶,由4個(gè)30-40kDa的亞基組 GAPDH蛋白幾乎在所有組織中都高水平表達(dá),廣泛用作Western blot蛋白質(zhì)標(biāo)準(zhǔn)化的內(nèi)參,是很好的內(nèi)參抗體。
GAPDH 作為管家基因在同種細(xì)胞或者組織中的蛋白質(zhì)表達(dá)量一般是恒定的。在實(shí)驗(yàn)中,可能存在總蛋白濃度測(cè)定不準(zhǔn)確;或者蛋白質(zhì)樣品在電泳前上樣時(shí)產(chǎn)生的樣品間的操作誤差;這些誤差需要通過(guò)測(cè)定每個(gè)樣品中實(shí)際轉(zhuǎn)到膜上的GAPDH的含量來(lái)進(jìn)行校正,所以一般的western實(shí)驗(yàn)都需要進(jìn)行內(nèi)參設(shè)置。具體校正的方法就是將每個(gè)樣品測(cè)得的目的蛋白含量與本樣品的GAPDH含量相除,得到每個(gè)樣品目的蛋白的相對(duì)含量。然后才進(jìn)行樣品與樣品之間的比較。
 
產(chǎn)品圖片 Tissue/cell: human lung carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-GAPDH Polyclonal Antibody, Unconjugated(bs-0755R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Tissue/cell: rat kidney carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-GAPDH Polyclonal Antibody, Unconjugated(bs-0755R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Tissue/cell: A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (GAPDH) polyclonal Antibody, Unconjugated (bs-0755R) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.Tissue/cell: A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (GAPDH) polyclonal Antibody, Unconjugated (bs-0755R) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.Blank control (Black line): U87MG (Black).
Primary Antibody (green line): Rabbit Anti-GAPDH antibody (bs-0755R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF647
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 10,000 events was performed.

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