重组
RabMAb

Anti-Bad抗体[Y208] (ab32445)

概述

  • 产品名称
    Anti-Bad抗体[Y208]
    参阅全部 Bad 一抗
  • 描述
    兔单克隆抗体[Y208] to Bad
  • 特异性
    The antibody does not cross-react with other Bcl-2 members.
  • 经测试应用
    适用于: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Dog, Human
  • 免疫原

    A synthetic peptide corresponding to residues in the N-term of human Bad.

  • 阳性对照
    • Hela cell lysate, human lymphoma.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

     

    Alternative versions available:

    Anti-Bad antibody (Biotin) [Y208] (ab199754)

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

性能

应用

Our Abpromise guarantee covers the use of ab32445 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 1/2000 - 1/5000. Detects a band of approximately 23 kDa (predicted molecular weight: 18 kDa).
IHC-P Use at an assay dependent concentration.
ICC/IF 1/250 - 1/500.
Flow Cyt 1/20 - 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/100.

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

Anti-Bad antibody [Y208] 图像

  • Immunocytochemical analysis of 2% paraformaldehyde-fixed,0.1% Triton-X100 permeabilized RAW 264.7 labeling Bad with ab32445 at 1/250dilution,followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 568) secondary antibody at 1/1000 dilution.

  • Immunohistochemical staining of paraffin-embedded human lymphoma using ab32445 at 1/100 dilution.
  • Anti-Bad antibody [Y208] (ab32445) at 1/5000 dilution + Hela cell lysate

    Predicted band size : 18 kDa
    Observed band size : 23 kDa (why is the actual band size different from the predicted?)
  • Overlay histogram showing MCF-7 cells stained with ab32445 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32445, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in MCF-7 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

Anti-Bad antibody [Y208] (ab32445)参考文献

This product has been referenced in:
  • Li Y  et al. IL-7 Receptor Mutations and Steroid Resistance in Pediatric T cell Acute Lymphoblastic Leukemia: A Genome Sequencing Study. PLoS Med 13:e1002200 (2016). WB ; Human . Read more (PubMed: 27997540) »
  • Yang Z  et al. Phosphorylation and inactivation of PTEN at residues Ser380/Thr382/383 induced by Helicobacter pylori promotes gastric epithelial cell survival through PI3K/Akt pathway. Oncotarget 6:31916-26 (2015). IHC-P ; Human . Read more (PubMed: 26376616) »

See all 18 Publications for this product

Product Wall

ab33906 lot YH090207C - 0.3260 mg/ml

ab33862 lot YH082410D - 0.6380 mg/ml

ab32429 lot YG080503 - 0.0790 mg/ml

ab40660 lot YI070802CS - 0.1030 mg/ml

ab32445 lot YI072804DS - 0.0250 mg/ml

Application
Immunocytochemistry
Sample
Mouse Cultured Cells (RAW 264.7)
Specification
RAW 264.7
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton-X100 in 2% BSA for 15min
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Username

Dr. Mahesh Shivananjappa

Verified customer

提交于 May 19 2012

Thank you for contacting us.
There is a wide range of secondaries suitable to detect either ab32445 or ab33862 in Flow Cytometry. I have conducted a search through our Secondaries search tool in the Abcam Website and found more than 100 matches, d...

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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:

- Return in the original Abcam box with...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Glioblastoma neurospheres cells (primary))
Loading amount
30 µg
Specification
Glioblastoma neurospheres cells (primary)
Gel Running Conditions
Reduced Denaturing (12.5% gel)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Dec 06 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Platelets)
Loading amount
20 µg
Specification
Platelets
Treatment
Calcium Ionophore for 1 h
Gel Running Conditions
Reduced Denaturing (4-20%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dr. Mahesh Shivananjappa

Verified customer

提交于 Nov 14 2011

Thank you for contacting us. I can confirm that this antibody does recongize the phosphorylated and unphosphorylated form of BAD. Indeed, there is no phosphorylation site on the immunogen sequences used for this antibody. I hope this inf...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (Mononuclear Cell Preparations)
Specification
Mononuclear Cell Preparations
Fixation
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton-X100 in 2% BSA for 15min
Gating Strategy
Monocytes Lymphocytes
Username

Dr. Mahesh Shivananjappa

Verified customer

提交于 Mar 30 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (PMN)
Specification
PMN
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton-X100 in 2% BSA for 30min
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Dec 20 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (293T)
Total protein in input
200 µg
Specification
293T
Immuno-precipitation step
Protein A
Username

Abcam user community

Verified customer

提交于 Dec 31 2007

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

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