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Rabbit Anti-GAPDH (Loading Control) antibody (bs-10900R)
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說明書: 50ul  100ul  200ul
50ul/680.00元
100ul/980.00元
200ul/1780.00元
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產品編號 bs-10900R
英文名稱 GAPDH (Loading Control)
中文名稱 3-磷酸甘油醛脫氫酶(內參)抗體
別    名 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.  
產品類型 內參抗體 
研究領域 腫瘤  細胞生物  免疫學  信號轉導  新陳代謝  
抗體來源 Rabbit
克隆類型 Polyclonal
交叉反應 Human, Mouse, Rat, 
產品應用 WB=1:5000-10000 IHC-P=1:100-500 IHC-F=1:100-500 ICC=1:100-500 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 38kDa
細胞定位 細胞核 細胞漿 細胞膜 
性    狀 Liquid
濃    度 1mg/ml
免 疫 原 Recombinant human GAPDH full length protein: 
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. 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: 24383 Rat

Entrez Gene: 685186 Rat

Entrez Gene: 317743 Zebrafish

Omim: 138400 Human

SwissProt: P00356 Chicken

SwissProt: P04406 Human

SwissProt: P16858 Mouse

SwissProt: P04797 Rat

SwissProt: Q5XJ10 Zebrafish



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

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。
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的亞基組成.
產品圖片
Sample:
293T(Human) Cell Lysate at 30 ug
Primary: Anti-GAPDH (bs-10900R) 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: 293T(human) cell lysate at 30ug;
Primary:
Lane1: Anti-GAPDH (bs-10900R) at 1/2000 dilution
Lane2: Anti-GAPDH (bs-10900R) at 1/10000 dilution
Lane3: Anti-GAPDH (bs-10900R) at 1/40000 dilution
Lane4: Anti-GAPDH (bs-10900R) at 1/80000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 38 kD
Observed band size: 38kD
Sample:
H9C2(Rat) Cell Lysate at 30 ug
Primary: Anti-GAPDH (bs-10900R) at 1/2000 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 (bs-10900R) 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:
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 (bs-10900R) 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:
Lung (Mouse) Lysate at 40 ug
Spleen (Mouse) Lysate at 40 ug
Thymus (Mouse) Lysate at 40 ug
Primary: Anti- GAPDH-Loading Control (bs-10900R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 38 kD
Observed band size: 35 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 (bs-10900R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 38 kD
Observed band size: 36 kD
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 (bs-10900R) at 1:2000 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse brain tissue); 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 Contro) Polyclonal Antibody, Unconjugated (bs-10900R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Human colon 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) Polyclonal Antibody, Unconjugated (bs-10900R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Human glioma); 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 (bs-10900R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Human liver cancer); 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 (bs-10900R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
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