bsm-52020R
[Primary Antibody]
Bad (5D4) Monoclonal Antibody
DATASHEET
Host: Rabbit
Target Protein: Bad
Clonality: Monoclonal
Isotype: IgG
Entrez Gene: 572
Swiss Prot: Q92934
Source: Recombinant human Bad protein, full length.
Purification: Purified by Protein A.
Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage: Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Background:
Bad is a member of the Bcl2 family and acts to promote apoptosis by forming heterodimers with the survival proteins Bcl2 and BclxL, thus preventing them from binding with BAX. Bad is found on the outer mitochondrial membrane and, once phosphorylated in response to growth stimuli, translocates to the cytoplasm. The phosphorylation status of Bad represents a key checkpoint for death or cell survival. JNK-induced phosphorylation of BAD serine 128 promotes the apoptotic role of Bad by opposing the inhibitory effect of growth factor on Bad-mediated apoptosis. Cdc2-induced phosphorylation of Bad serine 128 has an inhibitory effect on its interaction with 14-3-3 proteins. The latter interaction is critical for Bad phosphorylation at serine 155, a site within the BH3 domain that leads to the release of BclxL and the promotion of cell survival. Alternative splicing of this gene results in two transcript variants which encode the same isoform.
Size: 100ul
Concentration: 1ug/ul
Cross Reactive Species:
Human
Mouse
Rat
For research use only. Not intended for diagnostic or therapeutic use.
VALIDATION IMAGES
Lane 1: MCF-7 lysates probed with Bad (5D4) Monoclonal Antibody (bsm-52020R) at 1:500.
Paraformaldehyde-fixed, paraffin embedded; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer at 37°C for 20min; Antibody incubation with Bad (5D4) Monoclonal Antibody,(bsm-52020R) at 1:50 overnight at 4°C, followed by a conjugated secondary and DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat lung; 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 Bad (5D4) Monoclonal Antibody, Unconjugated (bsm-52020R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat skin; 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 Bad (5D4) Monoclonal Antibody, Unconjugated (bsm-52020R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse lung; 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 Bad (5D4) Monoclonal Antibody, Unconjugated (bsm-52020R) at 1:200 overnight at 4°C, 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 Bad (5D4) Monoclonal Antibody, Unconjugated (bsm-52020R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human stomach; 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 Bad (5D4) Monoclonal Antibody, Unconjugated (bsm-52020R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human breast 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 Bad (5D4) Monoclonal Antibody, Unconjugated (bsm-52020R) at 1:200 overnight at 4°C, 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 Bad (5D4) Monoclonal Antibody, Unconjugated (bsm-52020R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human pancreatic 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 Bad (5D4) Monoclonal Antibody, Unconjugated (bsm-52020R) at 1:200 overnight at 4°C, DAB staining.
Lane 1: Human Jurkat cell lysates; Lane 2: Human A549 cell lysates probed with Bad (5D4) Monoclonal Antibody, Unconjugated (bsm-52020R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.