bsm-33033M [Primary Antibody]
Mouse  Anti-GAPDH-Loading Control  Monoclonal Antibody
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Host: Mouse

Target Protein: GAPDH-Loading Control

IR: Immunogen Range:

Clonality: Monoclonal

Isotype: IgG

Entrez Gene: 2597

Swiss Prot: P04406

Source: Recombinded Human GAPDH : 

Purification: affinity purified by Protein G

Storage: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background: 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.

Size: 50ul

Concentration: 1mg/ml

Applications: WB=1:10000-1:100000,IHC-P=1:100-500,ICC=1:100

Cross Reactive Species: Human,Mouse,Rat,Pig,Hamster (predicted: Rabbit,Sheep,Chicken,Dog,Monkey)

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES
Sample: Lane1: Skin (Mouse) Lysate at 40 ug Lane2: Testis (Mouse) Lysate at 40 ug Lane3: Adrenal gland (Mouse) Lysate at 40 ug Primary: Anti-GAPDH (bsm-33033M) at 1/1 000 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution Predicted band size: 38 kDa Observed band size: 38 kDa
Sample: Lane 1: Rat Cerebrum tissue lysates Lane 2: Rat Heart tissue lysates Lane 3: Human A549 cell lysates Lane 4: Human Jurkat cell lysates Lane 5: Human Huvec cell lysates Lane 6: Human Hela cell lysates Lane 7: Human U2os cell lysates Lane 8: Human HepG2 cell lysates Primary: Anti- GAPDH (bsm-33033M) at 1/50000 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution Predicted band size: 38 kDa Observed band size: 38 kDa
Sample: Lane 1~4：Cerebrum (Rat) Lysate at 40 ug Lane 5~8：Hela (Human) Lysate at 40 ug Primary: Anti-GAPDH (bsm-33033M) at 1/2000~1/20000 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution Predicted band size: 38 kDa Observed band size: 38 kDa
Sample: Lane 1: Human HEK293 cell lysates Lane 2: Human Hela cell lysates Lane 3: Human SH-SY5Y cell lysates Lane 4: Mouse NIH/3T3 cell lysates Lane 5: Hamster CHO cell lysates Lane 6: Rat Brain tissue lysates Primary: Anti-GAPDH (bsm-33033M) at 1/200000 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution Predicted band size: 38 kDa Observed band size: 38 kDa
Paraformaldehyde-fixed, paraffin embedded (mouse brain); 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) Monoclonal Antibody, Unconjugated (ascites of bsm-33033M-4E8) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat spleen); 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) Monoclonal Antibody, Unconjugated (ascites of bsm-33033M-4E8) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat brain); 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) Monoclonal Antibody, Unconjugated (ascites of bsm-33033M-4E8) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse brain); 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) Monoclonal Antibody, Unconjugated (ascites of bsm-33033M-4E8) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat brain); 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) Monoclonal Antibody, Unconjugated (ascites of bsm-33033M-4E8) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat spleen); 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) Monoclonal Antibody, Unconjugated (ascites of bsm-33033M-4E8) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human gastric); 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) Monoclonal Antibody, Unconjugated (bsm-33033M ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) 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-Loading Control) Monoclonal Antibody, Unconjugated (bsm-33033M ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat brain); 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) Monoclonal Antibody, Unconjugated (bsm-33033M ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human gastric 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) Monoclonal Antibody, Unconjugated (bsm-33033M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human gastric); 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) Monoclonal Antibody, Unconjugated (bsm-33033M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) 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-Loading Control) Monoclonal Antibody, Unconjugated (bsm-33033M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat brain); 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) Monoclonal Antibody, Unconjugated (bsm-33033M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse brain); 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) Monoclonal Antibody, Unconjugated (bsm-33033M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Tissue/cell:Hela 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-Loading Control) monoclonal Antibody, Unconjugated (bsm-33033M) 1:100, 90 minutes at 37°C; followed by a CY3 conjugated Goat Anti-Mouse IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
 
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