Phospho-BAD (Ser112)[Ser75] Antibody - #AF7427

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Product: Phospho-BAD (Ser112)[Ser75] Antibody
Catalog: AF7427
Description: Rabbit polyclonal antibody to Phospho-BAD (Ser112)[Ser75]
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Bovine, Horse, Sheep, Dog
Mol.Wt.: 23kDa; 18kD(Calculated).
Uniprot: Q92934
RRID: AB_2843867

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Bovine(90%), Horse(92%), Sheep(91%), Dog(91%)
Clonality:
Polyclonal
Specificity:
Phospho-BAD (Ser112) Antibody detects endogenous levels of BAD only when phosphorylated at Ser75, which site historically referenced as Ser112.
RRID:
AB_2843867
Cite Format: Affinity Biosciences Cat# AF7427, RRID:AB_2843867.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

AI325008; BAD; BAD_HUMAN; 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 Death Promoter; Bcl-2-binding component 6; Bcl-2-like protein 8; Bcl-XL/Bcl-2-associated death promoter; Bcl2 antagonist of cell death; BCL2 antagonist of cell death protein; BCL2 associated agonist of cell death; Bcl2 Associated Death Promoter; BCL2 binding component 6; BCL2 binding protein; Bcl2 Like 8 Protein; Bcl2-L-8; BCL2L8; Proapoptotic BH3 Only Protein;

Immunogens

Immunogen:
Uniprot:

Q92934(BAD_HUMAN) >>Visit HPA database.

Gene(ID):
BAD(572)
Expression:
Q92934 BAD_HUMAN:

Expressed in a wide variety of tissues.

Sequence:
MFQIPEFEPSEQEDSSSAERGLGPSPAGDGPSGSGKHHRQAPGLLWDASHQQEQPTSSSHHGGAGAVEIRSRHSSYPAGTEDDEGMGEEPSPFRGRSRSAPPNLWAAQRYGRELRRMSDEFVDSFKKGLPRPKSAGTATQMRQSSSWTRVFQSWWDRNLGRGSSAPSQ

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Horse
92
Sheep
91
Dog
91
Bovine
90
Pig
0
Xenopus
0
Zebrafish
0
Chicken
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q92934 As Substrate

Site PTM Type Enzyme
M1 Acetylation
S25 Phosphorylation
S32 Phosphorylation
S34 Phosphorylation
K36 Acetylation
S71 Phosphorylation
S74 Phosphorylation Q13153 (PAK1)
S75 Phosphorylation Q9NQU5 (PAK6) , Q15349 (RPS6KA2) , Q9P1W9 (PIM2) , P31749 (AKT1) , P22612 (PRKACG) , P28482 (MAPK1) , P27361 (MAPK3) , O75582 (RPS6KA5) , Q04759 (PRKCQ) , Q13153 (PAK1) , P04049 (RAF1) , P41743 (PRKCI) , Q15418 (RPS6KA1) , P45983 (MAPK8) , Q02156 (PRKCE) , Q86V86 (PIM3) , Q15139 (PRKD1) , P17612 (PRKACA) , P22694 (PRKACB) , P51812 (RPS6KA3) , P11309 (PIM1) , O96013 (PAK4) , P15056 (BRAF) , P10398 (ARAF)
Y76 Phosphorylation
T80 Phosphorylation
S91 Phosphorylation P06493 (CDK1) , P45983 (MAPK8)
R94 Methylation
R96 Methylation
S97 Phosphorylation
S99 Phosphorylation Q9Y243 (AKT3) , P51812 (RPS6KA3) , Q04759 (PRKCQ) , Q13153 (PAK1) , P31751 (AKT2) , Q15139 (PRKD1) , P41743 (PRKCI) , P31749 (AKT1) , P15056 (BRAF) , P04049 (RAF1) , Q15418 (RPS6KA1) , Q86V86 (PIM3) , P23443 (RPS6KB1) , P17612 (PRKACA) , Q9P1W9 (PIM2) , Q02156 (PRKCE) , P11309 (PIM1) , P10398 (ARAF) , P22612 (PRKACG) , P22694 (PRKACB)
Y110 Phosphorylation
S118 Phosphorylation Q15139 (PRKD1) , Q02156 (PRKCE) , P31751 (AKT2) , Q13153 (PAK1) , Q86V86 (PIM3) , P17612 (PRKACA) , Q9P1W9 (PIM2) , P15056 (BRAF) , Q9Y243 (AKT3) , Q15349 (RPS6KA2) , P31749 (AKT1) , Q13976 (PRKG1) , P11309 (PIM1) , P10398 (ARAF) , P04049 (RAF1) , P22612 (PRKACG) , P22694 (PRKACB) , P41743 (PRKCI) , Q15418 (RPS6KA1)
S124 Phosphorylation
S128 Phosphorylation
S134 Phosphorylation Q13153 (PAK1) , P15056 (BRAF) , Q13555 (CAMK2G)
T137 Phosphorylation
T139 Phosphorylation
S146 Phosphorylation
S153 Phosphorylation P17612 (PRKACA) , O75582 (RPS6KA5) , Q15418 (RPS6KA1)
R161 Methylation

Research Backgrounds

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 (By similarity). Appears to act as a link between growth factor receptor signaling and the apoptotic pathways.

PTMs:

Phosphorylated on one or more of Ser-75, Ser-99, Ser-118 and Ser-134 in response to survival stimuli, which blocks its pro-apoptotic activity. Phosphorylation on Ser-99 or Ser-75 promotes heterodimerization with 14-3-3 proteins. This interaction then facilitates the phosphorylation at Ser-118, a site within the BH3 motif, leading to the release of Bcl-X(L) and the promotion of cell 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 blocked by the apoptotic C-terminus cleavage product of PKN2 generated by caspases-3 activity during apoptosis.

Methylation at Arg-94 and Arg-96 by PRMT1 inhibits Akt-mediated phosphorylation at Ser-99.

Subcellular Location:

Mitochondrion outer membrane. Cytoplasm.
Note: Colocalizes with HIF3A in the cytoplasm (By similarity). Upon phosphorylation, locates to the cytoplasm.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed in a wide variety of tissues.

Subunit Structure:

Forms heterodimers with the anti-apoptotic proteins, Bcl-X(L), Bcl-2 and Bcl-W. Also binds protein S100A10 (By similarity). The Ser-75/Ser-99 phosphorylated form binds 14-3-3 proteins (By similarity). Interacts with AKT1 and PIM3. Interacts (via BH3 domain) with NOL3 (via CARD domain); preventing the association of BAD with BCL2 (By similarity). Interacts with HIF3A (via C-terminus domain); the interaction reduces the binding between BAD and BAX (By similarity). Interacts with GIMAP3/IAN4 and GIMAP5/IAN5.

Family&Domains:

Intact BH3 motif is required by BIK, BID, BAK, BAD and BAX for their pro-apoptotic activity and for their interaction with anti-apoptotic members of the Bcl-2 family.

Belongs to the Bcl-2 family.

Research Fields

· Cellular Processes > Transport and catabolism > Autophagy - animal.   (View pathway)

· Cellular Processes > Cell growth and death > Apoptosis.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.   (View pathway)

· Environmental Information Processing > Signal transduction > ErbB signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Ras signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > cGMP-PKG signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > cAMP signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.

· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.

· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.

· Human Diseases > Neurodegenerative diseases > Alzheimer's disease.

· Human Diseases > Neurodegenerative diseases > Amyotrophic lateral sclerosis (ALS).

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

· Human Diseases > Infectious diseases: Viral > Hepatitis C.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Specific types > Colorectal cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Renal cell carcinoma.   (View pathway)

· Human Diseases > Cancers: Specific types > Pancreatic cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Endometrial cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Prostate cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Melanoma.   (View pathway)

· Human Diseases > Cancers: Specific types > Chronic myeloid leukemia.   (View pathway)

· Human Diseases > Cancers: Specific types > Acute myeloid leukemia.   (View pathway)

· Human Diseases > Cancers: Specific types > Non-small cell lung cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma.   (View pathway)

· Organismal Systems > Nervous system > Neurotrophin signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Insulin signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Thyroid hormone signaling pathway.   (View pathway)

References

1). Metal-coordinated nanomedicine for combined tumor therapy by inducing paraptosis and apoptosis. Journal of Controlled Release, 2021 (PubMed: 34146614) [IF=10.8]

Application: WB    Species: Mouse    Sample: 4T1 cells

Fig. 3. COMBO induced cell paraptosis and apoptosis. (A) Western blot analysis of the expression of Bad and cleaved caspase-3 by 4T1 cells after treatment with CODox, morusin and COMBO. Quantitative analysis of the fold changes of (B) Bad and (C) cleaved caspase-3 accumulated in 4T1 cells after treatment with CODox, morusin and COMBO (n = 2). (D) Western blot analysis of the expression of Alix, IRE1α, CHOP and p-eIF2α by 4T1 cells after treatment with CODox, morusin and COMBO. Quantitative analysis of the fold changes of (E) Alix, (F) IRE1α, (G) CHOP and (H) p-eIF2α accumulated in 4T1 cells after treatment with CODox, morusin and COMBO (n = 2). The cells were incubated with CODox, morusin or COMBO at the equivalent concentrations of DOX (0.6 mg/L), morusin (30 mg/L) and Cu2+ (1.2 mg/L). *P < 0.05, **P < 0.01 and ***P < 0.005 were tested via a Student’s t-test.
2). Cholesterol regulates cell proliferation and apoptosis of colorectal cancer by modulating miR-33a-PIM3 pathway. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2019 (PubMed: 30827510) [IF=3.1]

Application: WB    Species: human    Sample: CRC cells

Fig. 3.| Mechanism of miR-33a-PIM3 pathway in the role of cholesterol in CRC cells. (A) Expression of PIM3 mRNA was not changed in cholesterol treatment for 48h. (B) The changes of miR-33a after cells were transfected with miR-33a mimics or inhibitor for 24h. (C) PIM3 mRNA of cells did not show obviously difference after transfected with miR-33a mimics or inhibitor for 24h (D) PIM3 was proved to be the target gene of miR-33a by dual luciferase reporter assay. (E) The difference of expression of cell cycle and apoptotic related protein in miR-33a-PIM3 pathway between CON and CH.(F) The expression difference of cell cycle and apoptotic related protein in miR-33a-PIM3 pathway after cells were transfected with miR-33a mimics or inhibitor for 48h.
3). Methyl-β-cyclodextrin potentiates the BITC-induced anti-cancer effect through modulation of the Akt phosphorylation in human colorectal cancer cells. BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 2018 (PubMed: 30200817) [IF=1.6]

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