重组Anti-Bcl-2抗体[E17] (ab32124)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E17] to Bcl-2
- Suitable for: WB, IHC-P, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-Bcl-2抗体[E17]
参阅全部 Bcl-2 一抗 -
描述
兔单克隆抗体[E17] to Bcl-2 -
宿主
Rabbit -
特异性
Anti-Bcl-2 antibody [E17] (ab32124) recognises Bcl-2. It does not cross-react with other Bcl-2 family members.Bcl-2 has two isoforms, one is around 26kDa and the other is around 20kDa (PMID: 26009263, PMID: 10400666, PMID: 32377726). -
经测试应用
适用于: WB, IHC-P, IPmore details
不适用于: Flow Cyt or ICC/IF -
种属反应性
与反应: Human -
免疫原
Synthetic peptide within Human Bcl-2 aa 50-150. The exact sequence is proprietary.
Database link: P10415 -
阳性对照
- WB: MCF7, A431, Jurkat, HeLa, THP-1 and SH-SY5Y cell lysates; Wild type HAP1 whole cell lysate. IHC-P: Human B-cell lymphona and breast carcinoma tissues; Human UM xenografts; Human salivary glands; Human DLBCL U2932 cell line xenograft tissue. IP: Jurkat whole cell lysate (ab7899).
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常规说明
Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).
See other anti-rabbit secondary antibodies that can be used with this antibody.
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
解离常数(KD)
KD = 3.00 x 10 -11 M Learn more about KD -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
E17 -
同种型
IgG -
研究领域
相关产品
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Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
- Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
- Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
- Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
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Isotype control
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Positive Controls
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32124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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WB | (7) |
1/1000. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa).
We recommend using ab182858 for murine samples. |
IHC-P | (1) |
1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. We do not recommend rat and mouse samples with IHC. |
IP | (1) |
1/50.
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说明 |
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WB
1/1000. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa). We recommend using ab182858 for murine samples. |
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. We do not recommend rat and mouse samples with IHC. |
IP
1/50. |
靶标
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功能
Suppresses apoptosis in a variety of cell systems including factor-dependent lymphohematopoietic and neural cells. Regulates cell death by controlling the mitochondrial membrane permeability. Appears to function in a feedback loop system with caspases. Inhibits caspase activity either by preventing the release of cytochrome c from the mitochondria and/or by binding to the apoptosis-activating factor (APAF-1). May attenuate inflammation by impairing NLRP1-inflammasome activation, hence CASP1 activation and IL1B release (PubMed:17418785). -
组织特异性
Expressed in a variety of tissues. -
疾病相关
A chromosomal aberration involving BCL2 has been found in chronic lymphatic leukemia. Translocation t(14;18)(q32;q21) with immunoglobulin gene regions. BCL2 mutations found in non-Hodgkin lymphomas carrying the chromosomal translocation could be attributed to the Ig somatic hypermutation mechanism resulting in nucleotide transitions. -
序列相似性
Belongs to the Bcl-2 family. -
结构域
BH1 and BH2 domains are required for the interaction with BAX and for anti-apoptotic activity.
The BH4 motif is required for anti-apoptotic activity and for interaction with RAF1 and EGLN3.
The loop between motifs BH4 and BH3 is required for the interaction with NLRP1. -
翻译后修饰
Phosphorylation/dephosphorylation on Ser-70 regulates anti-apoptotic activity. Growth factor-stimulated phosphorylation on Ser-70 by PKC is required for the anti-apoptosis activity and occurs during the G2/M phase of the cell cycle. In the absence of growth factors, BCL2 appears to be phosphorylated by other protein kinases such as ERKs and stress-activated kinases. Phosphorylated by MAPK8/JNK1 at Thr-69, Ser-70 and Ser-87, wich stimulates starvation-induced autophagy. Dephosphorylated by protein phosphatase 2A (PP2A).
Proteolytically cleaved by caspases during apoptosis. The cleaved protein, lacking the BH4 motif, has pro-apoptotic activity, causes the release of cytochrome c into the cytosol promoting further caspase activity.
Monoubiquitinated by PARK2, leading to increase its stability. Ubiquitinated by SCF(FBXO10), leading to its degradation by the proteasome. -
细胞定位
Mitochondrion outer membrane. Nucleus membrane. Endoplasmic reticulum membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 596 Human
- Omim: 151430 Human
- SwissProt: P10415 Human
- Unigene: 150749 Human
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别名
- Apoptosis regulator Bcl 2 antibody
- Apoptosis regulator Bcl-2 antibody
- Apoptosis regulator Bcl2 antibody
see all
图片
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (ab32124)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human B-cell lymphoma tissue labelling Bcl-2 with purified ab32124 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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All lanes : Anti-Bcl-2 antibody [E17] (ab32124) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : BCL2 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 26 kDa
Observed band size: 26 kDaLanes 1- 2: Merged signal (red and green). Green - ab32124 observed at 26 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32124 was shown to react with BCL2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255364 (knockout cell lysate ab263752) was used. Wild-type HeLa and BCL2 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. ab32124 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (ab32124)This image is courtesy of an anonymous Abreview.
Formaldehyde-fixed, paraffin-embedded human DLBCL U2932 cell line xenograft tissue stained for Bcl-2 using ab32124 at 1/200 dilution in immunohistochemical analysis, followed by Goat anti-Rabbit IgG Alexa Fluor® 555.
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All lanes : Anti-Bcl-2 antibody [E17] (ab32124)
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : BCL2 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : THP-1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 26 kDaLanes 1 - 4: Merged signal (red and green). Green - ab32124 observed at 26 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32124 was shown to specifically react with BCL2 when BCL2 knockout samples were used. Wild-type and BCL2 knockout samples were subjected to SDS-PAGE. Ab32124 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging. 3% milk used as blocking agent.
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ab32124 (purified) at 1/30 immunoprecipitating Bcl-2 in Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (ab32124)Image from Némati F et al., PLoS One. 2014;9(1):e80836. Fig 2.; doi: 10.1371/journal.pone.0080836. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Bcl-2 expression determined by immunohistochemical analyses of the 4 human UM xenografts (between 3 to 5 tumors have been studied per condition).
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All lanes : Anti-Bcl-2 antibody [E17] (ab32124) at 1/1000 dilution (puriifed)
Lane 1 : MCF7 (human breast adenocarcinoma cell line) cell lysate
Lane 2 : A431 (human epidermoid carcinoma cell line) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 26 kDa
Observed band size: 26 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (ab32124)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human B cell lymphoma tissue labelling Bcl-2 with unpurified ab32124. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.
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All lanes : Anti-Bcl-2 antibody [E17] (ab32124) at 1/200 dilution
Lane 1 : MCF7 (human breast adenocarcinoma cell line) cell lysate
Lane 2 : SK-BR-3 (human mammary gland adenocarcinoma cell line) cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/50000 dilution
Predicted band size: 26 kDa
Observed band size: 26 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
MCF-7 cells express Bcl-2, while SK-BR-3 cells do not express Bcl-2 (PMID: 18430249)
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All lanes : Anti-Bcl-2 antibody [E17] (ab32124) at 1/10000 dilution (purified)
Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 3 : SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 26 kDa
Observed band size: 26 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Anti-Bcl-2 antibody [E17] (ab32124) at 1/1000 dilution (unpurified) + Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
Predicted band size: 26 kDa
Observed band size: 26 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (ab32124)Image from Szyszko EA et al., Arthritis Res Ther. 2011 Jan 7;13(1):R2. Fig 5.; doi:10.1186/ar3220; 7 January 2011, Arthritis Research & Therapy 2011, 13:R2
Immunohistochemical analysis of Human salivary glands taken from patients with primary Sjögren's syndrome, staining Bcl-2 with unpurified ab32124.
Antigen retrieval was performed via heat mediation in a citrate buffer (pH 6). Sections were blocked using 2% BSA, 10% normal serum and permeabilized with 0.5% Triton X-100. Samples were incubated with primary antibody (1/100) for one hour at room temperature. An Alexa Fluor® 594-conjugated anti-rabbit IgG was used as the secondary antibody.
N.B. Panels B and D are higher magnifications of panels A and C, respectively. -
Equilibrium disassociation constant (KD)
Click here to learn more about KD -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (ab32124)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Bcl-2 with unpurified ab32124 at 1/200 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
文献 (867)
ab32124 被引用在 867 文献中.
- Bi Y et al. Long noncoding RNA HNF1A-AS1 regulates proliferation and apoptosis of glioma through activation of the JNK signaling pathway via miR-363-3p/MAP2K4. J Cell Physiol 236:1068-1082 (2021). PubMed: 32779194
- Jia G et al. Neural stem cell-conditioned medium ameliorates Aß25-35-induced damage in SH-SY5Y cells by protecting mitochondrial function. Bosn J Basic Med Sci 21:179-186 (2021). PubMed: 32156251
- Liu F et al. Upregulation of circ_0000142 promotes multiple myeloma progression by adsorbing miR-610 and upregulating AKT3 expression. J Biochem 169:327-336 (2021). PubMed: 32970816
- Yao W et al. TNK2-AS1 upregulated by YY1 boosts the course of osteosarcoma through targeting miR-4319/WDR1. Cancer Sci 112:893-905 (2021). PubMed: 33164271
- Wang YY et al. Nogo-A aggravates oxidative damage in oligodendrocytes. Neural Regen Res 16:179-185 (2021). PubMed: 32788474