[KO验证抗体] Rabbit Anti-Phospho-Bad (Ser118) antibody

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Bad (phospho S118); Bad (phospho Ser118); p-Bad (S118);p- Bad (Ser118); p-Bad (phospho Ser118); BBC 2; BBC2; BBC6; Bcl 2 Antagonist of Cell Death; Bcl 2 Binding Component 6; BCL X / BCL 2 Binding Protein; BCL X Binding Protein; Bcl XL/Bcl 2 Associated Dea

Cat:

SL5216R

Species Reactivity：

Human,Mouse,Rat,(predicted: Dog,Pig,Cow,Horse,Rabbit,)

Immunogen：

KLH conjugated Synthesised phosphopeptide derived from human BAD around the phosphorylation site of Ser118:RM(p-S)DE

Format：

Liquid

Storage instructions：

Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Concentration：

1mg/ml

Clonality：

Polyclonal

Isotype：

IgG

Applications：

WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500IHC-F=1:100-500Flow-Cyt=2ug/TestIF=1:100-500（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.

Host：

Rabbit

Product Overview：

Sample: Hela(Human) Cell Lysate at 30 ugHela KO Bad (Human) Cell Lysate at 30 ugPrimary: Anti- Phospho-Bad (Ser118) (SL5216R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 22 kDObserved band size: 22 kDSample: Hela(Human) Cell Lysate at 30 ugPrimary: Anti-Phospho-Bad (Ser118) (SL5216R) at 1/500 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 22 kDObserved band size: 22 kDSample: Raw264.7(Mouse) Cell Lysate at 30 ugPrimary: Anti-Phospho-Bad (Ser118) (SL5216R) at 1/500 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 22 kDObserved band size: 22 kDParaformaldehyde-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 (Phospho-Bad (Ser118)) Polyclonal Antibody, Unconjugated (SL5216R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (Human brain 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 (Phospho-Bad (Ser118)) Polyclonal Antibody, Unconjugated (SL5216R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) 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 (Phospho-Bad (Ser118)) Polyclonal Antibody, Unconjugated (SL5216R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Blank control:Jurkat. Primary Antibody (green line): Rabbit Anti-Phospho-Bad (Ser118) antibody (SL5216R) Dilution: 2ug/Test; Secondary Antibody : Goat anti-rabbit IgG-FITCDilution: 0.5ug/Test. ProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST 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 20,000 events was performed.Blank control (Black line): Jurkat (Black). Primary Antibody (green line): Rabbit Anti-Bad(Ser118) antibody (SL5216R) Dilution: 3μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody (white blue line): Goat anti-rabbit IgG-PEDilution: 1μg /test. ProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 15 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 20,000 events was performed.

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Unit:

Price: $256.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
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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 SH3 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.

Function:
Promotes cell death. Successfully competes for the binding to Bcl-X(L), Bcl-2 and Bcl-W, thereby affecting the level of heterodimerization of these proteins with BAX. Can reverse the death repressor activity of Bcl-X(L), but not that of Bcl-2. Appears to act as a link between growth factor receptor signaling and the apoptotic pathways.

Subunit:
Forms heterodimers with the anti-apoptotic proteins, Bcl-X(L), Bcl-2 and Bcl-W. Also binds protein S100A10. The Ser-75/Ser-99 phosphorylated form binds 14-3-3 proteins. Interacts with AKT1 and PIM3.

Subcellular Location:
Mitochondrion outer membrane. Cytoplasm. Upon phosphorylation, locates to the cytoplasm.

Tissue Specificity:
Expressed in a wide variety of tissues.

Post-translational modifications:
Phosphorylated on one or more of Ser-75, Ser-99, Ser-118 andSer-134 in response to survival stimuli, which blocks itspro-apoptotic activity. Phosphorylation on Ser-99 or Ser-75promotes heterodimerization with 14-3-3 proteins. This interactionthen facilitates the phosphorylation at Ser-118, a site within theSH3 motif, leading to the release of Bcl-X(L) and the promotion ofcell survival. Ser-99 is the major site of AKT/PKB phosphorylation, Ser-118 the major site of protein kinase A (CAPK) phosphorylation. Phosphorylation at Ser-99 by PKB/AKT1 is almost completely blockedby the apoptotic SLCterminus cleavage product of PKN2 generated bycaspases-3 activity during apoptosis.
Methylation at Arg-94 and Arg-96 by PRMT1 inhibits Akt-mediated phosphorylation at Ser-99.

Similarity:
Belongs to the Bcl-2 family.

SWISS:
Q92934

Gene ID:
572

Database links:

Entrez Gene: 572 Human

Entrez Gene: 12015 Mouse

Entrez Gene: 64639 Rat

Omim: 603167 Human

SwissProt: Q92934 Human

SwissProt: Q61337 Mouse

SwissProt: O35147 Rat

Unigene: 370254 Human

Unigene: 4387 Mouse

Unigene: 36696 Rat

BAD是BCL2/BAX、BCL-XL/BAX异二聚体的负调节基因。BAD是BCL2/BCL-XL相关死亡促进因子，作为BCL2、 bCL-XL异二聚体伴分子而促进细胞凋亡。 Picture

Sample:
Hela(Human) Cell Lysate at 30 ug
Primary: Anti-Phospho-Bad (Ser118) (SL5216R) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 22 kD
Observed band size: 22 kD

Sample:
Raw264.7(Mouse) Cell Lysate at 30 ug
Primary: Anti-Phospho-Bad (Ser118) (SL5216R) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 22 kD
Observed band size: 22 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 (Phospho-Bad (Ser118)) Polyclonal Antibody, Unconjugated (SL5216R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Human brain 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 (Phospho-Bad (Ser118)) Polyclonal Antibody, Unconjugated (SL5216R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) 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 (Phospho-Bad (Ser118)) Polyclonal Antibody, Unconjugated (SL5216R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Blank control:Jurkat.
Primary Antibody (green line): Rabbit Anti-Phospho-Bad (Ser118) antibody (SL5216R)
Dilution: 2ug/Test;
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 0.5ug/Test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST 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 20,000 events was performed.

Blank control (Black line): Jurkat (Black).
Primary Antibody (green line): Rabbit Anti-Bad(Ser118) antibody (SL5216R)
Dilution: 3μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 15 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 20,000 events was performed.

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