概述

  • 产品名称
  • 描述
    兔多克隆抗体to Bad
  • 宿主
    Rabbit
  • 特异性
    This antibody specifically binds the Bad protein, but does not recognize Bad/Bcl2 heterodimers in Immunoprecipitation assays.
  • 经测试应用
    适用于: WB, IHC-P, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Sheep, Rabbit, Chicken, Hamster, Cow, Dog, Pig, Drosophila melanogaster, Monkey
  • 免疫原

    Synthetic peptide derived from the sequence of Mouse Bad, conjugated to KLH.

  • 阳性对照
    • WB: Mouse Raw 264 cell lysate, Rat brain tissue lysate IHC-P: Alzheimer diseased brain section IF/ICC: RAW246.7 cell line

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能
    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.
  • 组织特异性
    Expressed in a wide variety of tissues.
  • 序列相似性
    Belongs to the Bcl-2 family.
  • 结构域
    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.
  • 翻译后修饰
    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.
  • 细胞定位
    Mitochondrion outer membrane. Cytoplasm. Upon phosphorylation, locates to the cytoplasm.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

图片

  • 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 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

  • 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).
  • 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.

文献

This product has been referenced in:
  • Yan H  et al. NR4A1-induced increase in the sensitivity of a human gastric cancer line to TNFa-mediated apoptosis is associated with the inhibition of JNK/Parkin-dependent mitophagy. Int J Oncol 52:367-378 (2018). Read more (PubMed: 29207128) »
  • Yan H  et al. Yap regulates gastric cancer survival and migration via SIRT1/Mfn2/mitophagy. Oncol Rep 39:1671-1681 (2018). Read more (PubMed: 29436693) »

See all 4 Publications for this product

客户评价及客户问答

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Abcam guarantees this product to work in the species/application used in this Abreview.
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
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提交于 Sep 27 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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