bsm-0978M [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: 100009074

Swiss Prot: P46406

Source: Full length GAPDH protein purified from rabbit muscle: 

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=1:1000-5000,IHC-P=1:100-500,IHC-F=1:100-500,ICC=1:100,IF=1:100-500

Cross Reactive Species: Human,Mouse,Rat (predicted: Rabbit)

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

VALIDATION IMAGES
Sample: Placenta (Mouse) Lysate at 40 ug 293T Cell Lysate at 40 ug Primary: Anti-GAPDH(bsm-0978M)at 1/1000 dilution Secondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilution Predicted band size: 38kD Observed band size: 38kD
Sample: Bladder(Rat)Lysate at 40 ug Primary: Anti-GAPDH(bsm-0978M)at 1/300 dilution Secondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilution Predicted band size: 38kD Observed band size: 38kD
Sample:Liver(Mouse)Lysate at 40 ug Primary: Anti-GAPDH(bsm-0978M)at 1/300 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution Predicted band size: 38kD Observed band size: 38kD
Sample: A549(Human) Cell Lysate at 30 ug 293T(Human) Cell Lysate at 30 ug HepG2(Human) Cell Lysate at 30 ug Hela(Human) Cell Lysate at 30 ug Primary: Anti- GAPDH(3E12) (bsm-0978M) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution Predicted band size: 38 kD Observed band size: 38 kD
Sample: Lane 1: Hela (Human) Cell Lysate at 30 ug Lane 2: Huvec (Human) Cell Lysate at 30 ug Lane 3: A549 (Human) Cell Lysate at 30 ug Lane 4: MCF-7 (Human) Cell Lysate at 30 ug Primary: Anti-GAPDH (bsm-0978M) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution Predicted band size: 36 kD Observed band size: 36 kD
Sample: Cerebrum (Mouse) Lysate at 40 ug Primary: Anti-GAPDH(3E12)-Loading Control (bsm-0978M) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution Predicted band size: 38 kD Observed band size: 38 kD
Paraformaldehyde-fixed, paraffin embedded (human breast 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-0978M) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand 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)) Monoclonal Antibody, Unconjugated (bsm-0978M) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human 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-0978M) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) 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)) Monoclonal Antibody, Unconjugated (bsm-0978M) at 1:100 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-0978M) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) 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)) Monoclonal Antibody, Unconjugated (bsm-0978M) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) 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))Monoclonal Antibody, Unconjugated (bsm-0978M) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat heart); 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-0978M) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand 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 (bsm-0978M) at 1:100 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-0978M) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand 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 (bsm-0978M) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human Prostate Tumor); 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-0978M) at 1:100 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-0978M) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
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)) Monoclonal Antibody, Unconjugated (bsm-0978M) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) 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(3E12)-Loading Control) Polyclonal Antibody, Unconjugated (bsm-0978M) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Tissue/cell: HUVEC 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(3E12)-Loading Control) Polyclonal Antibody, Unconjugated (bsm-0978M) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0296G-FITC) at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
 
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