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  1. Link

    bad-antibody-ab90435.pdf

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Cell Biology Apoptosis Intracellular Bcl2 Family
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Anti-Bad antibody (ab90435)

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

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

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  • Datasheet
  • References (17)
  • Protocols

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

  • 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...
  • Purity

    Protein A purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Bcl2 Family
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Bcl 2 family
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Bcl 2 family
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • RAW 264.7 whole cell lysate (ab7187)
  • Recombinant Protein

    • Recombinant Human Bad protein (ab89848)

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.
  • Target information above from: UniProt accession Q92934 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • 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.

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Datasheets and documents

    • Datasheet
  • 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

    Show All Reviews Q&A
    Submit a review Submit a question

    1-2 of 2 Abreviews or Q&A

    Western blot abreview for Anti-Bad antibody

    Good
    Abreviews
    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

    Abcam community

    Verified customer

    Asked on Feb 09 2012

    Answer

    Thank you for contacting Abcam.

    Included in this email are the instructions for returning your item to Abcam. When you return the goods, please ensure that you follow the procedures as below:

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    Read More

    Abcam Scientific Support

    Answered on Feb 09 2012

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