Anti-Bad antibody (ab90435)
- Datasheet
- References (17)
- Protocols
Overview
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Product name
Anti-Bad antibody
See all Bad primary antibodies -
Description
Rabbit polyclonal to Bad -
Host species
Rabbit -
Specificity
This antibody specifically binds the Bad protein, but does not recognize Bad/Bcl2 heterodimers in Immunoprecipitation assays. -
Tested applications
Suitable for: WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Sheep, Rabbit, Chicken, Hamster, Cow, Dog, Pig, Drosophila melanogaster, Monkey -
Immunogen
Synthetic peptide derived from the sequence of Mouse Bad, conjugated to KLH.
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Positive control
- WB: Mouse Raw 264 cell lysate, Rat brain tissue lysate IHC-P: Alzheimer diseased brain section IF/ICC: RAW246.7 cell line
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
Preservative: 0.09% Sodium Azide
Constituents: 50% Glycerol, PBS, pH 7.2 -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab90435 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | Use a concentration of 1 µg/ml. Predicted molecular weight: 18 kDa. | |
IHC-P | 1/50. Antigen retrieval is not essential but may optimise staining. | |
ICC/IF | Use a concentration of 5 µg/ml. |
Target
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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. -
Tissue specificity
Expressed in a wide variety of tissues. -
Sequence similarities
Belongs to the Bcl-2 family. -
Domain
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. -
Post-translational
modificationsPhosphorylated 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. Ser-75 is phosphorylated by AKT/PKB, protein kinase A and PIM2. -
Cellular localization
Mitochondrion outer membrane. Cytoplasm. Upon phosphorylation, locates to the cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 483763 Dog
- Entrez Gene: 12015 Mouse
- Entrez Gene: 64639 Rat
- SwissProt: Q61337 Mouse
- SwissProt: O35147 Rat
- Unigene: 4387 Mouse
- Unigene: 36696 Rat
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Alternative names
- AI325008 antibody
- BAD antibody
- BAD_HUMAN antibody
see all
Images
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Anti-Bad antibody (ab90435) at 1 µg/ml + Mouse Raw 264 cell lysate
Developed using the ECL technique.
Predicted band size: 18 kDa -
All lanes : Anti-Bad antibody (ab90435) at 1/1000 dilution
Lanes 1 & 5 : Cow normal mammary epithelial cells (nMEC) treated with serum free media
Lanes 2 & 6 : Cow normal mammary epithelial cells (nMEC) treated with 1.0µM anisomycin
Lanes 3 & 7 : Cow normal mammary epithelial cells (nMEC) treated with 100ng/ml IGF-1
Lanes 4 & 8 : Cow normal mammary epithelial cells (nMEC) treated with anisomycin and IGF-1
Lysates/proteins at 40 µg per lane.
Secondary
All lanes : ECL Anti-Rabbit IgG, Horseradish Peroxidase Linked at 2500 cells
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 18 kDa
Observed band size: 18 kDa
Exposure time: 30 secondsTreatement for 30 minutes (lanes 1-4) or for 1 hour (lanes 5-8). 12.5% SDS-PAGE. Proteins collected in RIPA Buffer and transferred to PVDF (0.45 um).
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ab90435, at a 1/50 dilution, staining BAD in the neurons of paraffin embedded Alzheimer diseased brain tissue (right) by Immunohistochemistry. Control brain tissue (left).
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ICC/IF image of ab90435 stained RAW246.7 cells. The cells were 4% paraformaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab90435, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Protocols
Datasheets and documents
References
This product has been referenced in:
- Ma S & Dong Z Melatonin Attenuates Cardiac Reperfusion Stress by Improving OPA1-Related Mitochondrial Fusion in a Yap-Hippo Pathway-Dependent Manner. J Cardiovasc Pharmacol 73:27-39 (2019). Read more (PubMed: 30418242) »
- Zhou T et al. Mst1 inhibition attenuates non-alcoholic fatty liver disease via reversing Parkin-related mitophagy. Redox Biol 21:101120 (2019). Read more (PubMed: 30708325) »