Bad (Ser118) Polyclonal Antibody

$Paraformaldehyde-fixed, paraffin embedded rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 20min; Antibody incubation with Bad (Ser118) Polyclonal Antibody, Unconjugated (bs-5216R) at 1:200 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.$

Paraformaldehyde-fixed, paraffin embedded rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 20min; Antibody incubation with Bad (Ser118) Polyclonal Antibody, Unconjugated (bs-5216R) at 1:200 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.

$Paraformaldehyde-fixed, paraffin embedded human glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 20min; Antibody incubation with Bad (Ser118) Polyclonal Antibody, Unconjugated (bs-5216R) at 1:200 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.$

Paraformaldehyde-fixed, paraffin embedded human glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 20min; Antibody incubation with Bad (Ser118) Polyclonal Antibody, Unconjugated (bs-5216R) at 1:200 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.

Hela cell lysates probed with Phospho-Bad (Ser118) Polyclonal Antibody, Unconjugated (bs-5216R) at 1:500 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

Lane 1: Hela cell lysates; Lane 2: Hela KO Bad cell lysates probed with Bad (Ser118) Polyclonal Antibody, Unconjugated (bs-5216R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

$Lane 1: Mouse Raw264.7 Cells; 30ug loaded in each lane; Probed with Bad (Ser118) Polyclonal Antibody, unconjugated (bs-5216R) at 1:500 overnight at 4\u00b0C followed by a conjugated secondary antibody for 60 minutes at 37\u00b0C.$

Lane 1: Mouse Raw264.7 Cells; 30ug loaded in each lane; Probed with Bad (Ser118) Polyclonal Antibody, unconjugated (bs-5216R) at 1:500 overnight at 4\u00b0C followed by a conjugated secondary antibody for 60 minutes at 37\u00b0C.

Human Jurkat 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 Bad (Ser118) Polyclonal Antibody Unconjugated (bs-5216R; green) at 1:33 for 30 min at room temperature. The secondary antibody, Goat anti-rabbit IgG-PE, was used for 40 min at room temperature. Staining is compared to compared to unstained cells (dark blue), secondary only (light blue), and isotype control (orange).

$Jurkat cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Bad (Ser118) Polyclonal Antibody(bs-5216R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).\r\n$

Jurkat cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Bad (Ser118) Polyclonal Antibody(bs-5216R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).\r\n

$Paraformaldehyde-fixed, paraffin embedded (Human kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (Phospho-Bad (Ser118)) Polyclonal Antibody, Unconjugated (bs-5216R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.$

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\u00b0C for 30min; Antibody incubation with (Phospho-Bad (Ser118)) Polyclonal Antibody, Unconjugated (bs-5216R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Applications

WB
ELISA
FCM
IHC-P
IHC-F
IF(IHC-P)
IF(IHC-F)
IF(ICC)
ICC

Reactivity

Human
Mouse
Rat

Predicted Reactivity

Dog
Cow
Pig
Horse
Rabbit

Overview
Catalog #	bs-5216R
Product Name	Bad (Ser118) Polyclonal Antibody
Applications	WB, ELISA, FCM, IHC-P, IHC-F, IF(IHC-P), IF(IHC-F), IF(ICC), ICC
Specificity	KO-Validated
Reactivity	Human, Mouse, Rat
Predicted Reactivity	Dog, Cow, Pig, Horse, Rabbit
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic phosphopeptide derived from human BAD around the phosphorylation site of Ser118
Modification Site	Ser118
Clonality	Polyclonal
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	572
Swiss Prot	Q92934
Subcellular location	Cytoplasm, Cell membrane
Synonyms	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; mouse BAD Ser155, rat BAD Ser156
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.
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
FCM	1:20-100
IHC-P	1:200-400
IHC-F	1:100-500
IF(IHC-P)	1:50-200
IF(IHC-F)	1:50-200
IF(ICC)	1:50-200
ICC	1:100-500