Anti-Bad antibody (ab90435)

Reviews (1)Q&A (1)References (17)

Overview

  • 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.

  • 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

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
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

  • 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
    modifications

    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. 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

  • Database links

    • Alternative names

      • AI325008 antibody
      • BAD antibody
      • BAD_HUMAN antibody
      • BBC 2 antibody
      • BBC2 antibody
      • BBC6 antibody
      • Bcl 2 Antagonist of Cell Death antibody
      • Bcl 2 Binding Component 6 antibody
      • BCL X / BCL 2 Binding Protein antibody
      • BCL X Binding Protein antibody
      • Bcl XL/Bcl 2 Associated Death Promoter antibody
      • Bcl-2-binding component 6 antibody
      • Bcl-2-like protein 8 antibody
      • Bcl-XL/Bcl-2-associated death promoter antibody
      • Bcl2 antagonist of cell death antibody
      • BCL2 antagonist of cell death protein antibody
      • BCL2 associated agonist of cell death antibody
      • Bcl2 Associated Death Promoter antibody
      • BCL2 binding component 6 antibody
      • BCL2 binding protein antibody
      • Bcl2 Like 8 Protein antibody
      • Bcl2-L-8 antibody
      • BCL2L8 antibody
      • Proapoptotic BH3 Only Protein antibody
      see all

    Images

    • Western blot - Anti-Bad antibody (ab90435)
      Western blot - Anti-Bad antibody (ab90435)
      Anti-Bad antibody (ab90435) at 1 µg/ml + Mouse Raw 264 cell lysate

      Developed using the ECL technique.

      Predicted band size: 18 kDa

    • Western blot - Anti-Bad antibody (ab90435)
      Western blot - Anti-Bad antibody (ab90435)This image is courtesy of an anonymous Abreview
      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 seconds


      Treatement 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).

      See Abreview

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bad antibody (ab90435)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bad antibody (ab90435)
      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).
    • Immunocytochemistry/ Immunofluorescence - Anti-Bad antibody (ab90435)
      Immunocytochemistry/ Immunofluorescence - Anti-Bad antibody (ab90435)
      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.

    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) »
    See all 17 Publications for this product

    Publishing research using ab90435? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Western blot abreview for Anti-Bad antibody

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    40 µg
    Gel Running Conditions
    Reduced Denaturing (12.5% SDS-PAGE)
    Sample
    Cow Cell lysate - whole cell (Normal mammary epithelial cells (nMEC))
    Specification
    Normal mammary epithelial cells (nMEC)
    Treatment
    1.0 uM ANS +/- 100 ng/mL IGF-1
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 27 2013

    Question

    Ordered ab90435 by mistake and would like to return it.

    Read More
    Answer

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