| 中文名稱 | 3-磷酸甘油醛脫氫酶 |
| 別 名 | Glyceraldehyde-3-phosphate dehydrogenase. |
| 研究領域 | 細胞生物 免疫學 |
| 抗體來源 | Rabbit |
| 克隆類型 | Polyclonal |
| 交叉反應 | Human, Rat, (predicted: Mouse, ) |
| 產品應用 | 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） not yet tested in other applications. optimal dilutions/concentrations should be determined by the end user. |
| 分 子 量 | 37kDa |
| 細胞定位 | 細胞核 細胞漿 細胞膜 |
| 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 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）過程中的關鍵酶。除了在胞質中作為糖酵解的酶以外，有證據表明哺乳動物細胞中的GAPDH參與了多種胞內生化過程，包括膜融合（membrane fusion）、微管成束（microtubule bundling）、磷酸轉移酶（phosphotransferase）激活、核內RNA出核、DNA復制與DNA修復。一些生理因素，諸如低氧（hypoxia）和尿糖（diabetes），可以增加GAPDH在特定細胞中的表達。GAPDH存在于幾乎所有的組織中，以高水平持續表達。 GAPDH（甘油醛-3-磷酸脫氫酶）是參與糖酵解的一種關鍵酶，由4個30－40kDa的亞基組 GAPDH蛋白幾乎在所有組織中都高水平表達，廣泛用作Western blot蛋白質標準化的內參,是很好的內參抗體。 GAPDH 作為管家基因在同種細胞或者組織中的蛋白質表達量一般是恒定的。在實驗中，可能存在總蛋白濃度測定不準確；或者蛋白質樣品在電泳前上樣時產生的樣品間的操作誤差；這些誤差需要通過測定每個樣品中實際轉到膜上的GAPDH的含量來進行校正，所以一般的western實驗都需要進行內參設置。具體校正的方法就是將每個樣品測得的目的蛋白含量與本樣品的GAPDH含量相除，得到每個樣品目的蛋白的相對含量。然后才進行樣品與樣品之間的比較。 |
| 產品圖片 |  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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