重组Anti-Bad抗体[Y208] - BSA and Azide free (ab220116)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y208] to Bad - BSA and Azide free
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Bad抗体[Y208] - BSA and Azide free
参阅全部 Bad 一抗 -
描述
兔单克隆抗体[Y208] to Bad - BSA and Azide free -
宿主
Rabbit -
特异性
This antibody does not cross-react with other Bcl-2 members.
The mouse and rat recommendation is based on the IHC-P results. We do not guarantee WB for mouse and rat.
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经测试应用
适用于: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details -
种属反应性
与反应: Mouse, Rat, Human
预测可用于: Dog -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- IHC-P: Human colon, ovarian cancer, Mouse and Rat kidney tissue; ICC/IF: HeLa cells; Flow Cyt (intra): MCF7 cells. WB: HeLA and HepG2 whole cell lysate.
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常规说明
ab220116 is the carrier-free version of ab32445.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
存储溶液
pH: 7.20
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
Y208 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Biotin Anti-Bad antibody [Y208] (ab199754)
- PE Anti-Bad antibody [Y208] (ab305823)
- APC Anti-Bad antibody [Y208] (ab305824)
- HRP Anti-Bad antibody [Y208] (ab305825)
- Alexa Fluor® 488 Anti-Bad antibody [Y208] (ab309767)
- Alexa Fluor® 647 Anti-Bad antibody [Y208] (ab310139)
- Alexa Fluor® 594 Anti-Bad antibody [Y208] (ab310561)
- Alexa Fluor® 555 Anti-Bad antibody [Y208] (ab312090)
- Alexa Fluor® 568 Anti-Bad antibody [Y208] (ab312570)
- Anti-Bad antibody [Y208] (ab32445)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab220116于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 23 kDa (predicted molecular weight: 18 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
The mouse and rat recommendation is based on the IHC-P results. We do not guarantee WB for mouse and rat. |
说明 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 23 kDa (predicted molecular weight: 18 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. The mouse and rat recommendation is based on the IHC-P results. We do not guarantee WB for mouse and rat. |
靶标
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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
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数据库链接
- Entrez Gene: 483763 Dog
- Entrez Gene: 572 Human
- Entrez Gene: 12015 Mouse
- Entrez Gene: 64639 Rat
- Omim: 603167 Human
- SwissProt: Q92934 Human
- SwissProt: Q61337 Mouse
- SwissProt: O35147 Rat
see all -
别名
- AI325008 antibody
- BAD antibody
- BAD_HUMAN antibody
see all
图片
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All lanes : Anti-Bad antibody [Y208] (ab32445) at 1/2000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 18 kDa -
All lanes : Anti-Bad antibody [Y208] (ab32445) at 1/2000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : BAD knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 18 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab32445).
Lanes 1- 2: Merged signal (red and green). Green - ab32445 observed at 23 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32445 was shown to react with Bad in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264843 (knockout cell lysate ab256847) was used. Wild-type HeLa and BAD knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32445 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-Bad antibody [Y208] (ab32445) at 1/2000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : BAD knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : MCF7 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 18 kDaThis data was developed using the same antibody clone in a different buffer formulation (ab32445).
Lanes 1 - 4: Merged signal (red and green). Green - ab32445 observed at 23 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab32445 was shown to specifically recognise BAD in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when BAD knockout cells were examined. Wild-type and BAD knockout samples were subjected to SDS-PAGE. Ab32445 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.
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This data was developed using ab32445, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling Bad with purified ab32445 at 1/1000 dilution (0.14 µg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
This data was developed using ab32445, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Bad with purified ab32445 at 1/50 dilution (2.9 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488 ,ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control. -
This data was developed using ab32445, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian cancer tissue sections labeling Bad with purified ab32445 at 1/1000 dilution (0.14 µg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
This data was developed using ab32445, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labelling Bad with purified ab32445 at 1/20 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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This data was developed using ab32445, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling Bad with purified ab32445 at 1/1000 dilution (0.14 µg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
This data was developed using ab32445, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling Bad with purified ab32445 at 1/1000 dilution (0.14 µg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
实验方案
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (1)
ab220116 被引用在 1 文献中.
- Yu B et al. MicroRNA-124 suppresses growth and aggressiveness of osteosarcoma and inhibits TGF-ß-mediated AKT/GSK-3ß/SNAIL-1 signaling. Mol Med Rep 17:6736-6744 (2018). PubMed: 29488603