重组Anti-Bcl-2抗体[EPR17509] (ab182858)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17509] to Bcl-2
- Suitable for: Flow Cyt (Intra), WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-Bcl-2抗体[EPR17509]
参阅全部 Bcl-2 一抗 -
描述
兔单克隆抗体[EPR17509] to Bcl-2 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-Pmore details
不适用于: ICC/IF -
种属反应性
与反应: Mouse, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Human tonsil and thymus lysates; Jurkat, U-937, THP-1, HeLa, C2C12, WEHI -3 and NIH/3T3 whole cell lysates; Mouse brain, heart, kidney and spleen lysates; Human fetal kidney and fetal spleen lysates; Wild-type Hap1 cell lysate. IHC-P: Human tonsil tissue, Human endometrial cancer tissue, Mouse spleen tissue. Flow Cyt (intra): Jurkat cells.
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常规说明
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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
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
EPR17509 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab182858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) |
1/250.
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WB | (5) |
1/2000. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa).
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IHC-P | (7) |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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说明 |
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Flow Cyt (Intra)
1/250. |
WB
1/2000. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa). |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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
- Entrez Gene: 12043 Mouse
- Omim: 151430 Human
- SwissProt: P10415 Human
- SwissProt: P10417 Mouse
- Unigene: 150749 Human
- Unigene: 257460 Mouse
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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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All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/2000 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 - ab182858 observed at 26 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab182858 was shown to react with Bcl-2 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. ab182858 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] (ab182858)
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Bcl-2 with ab182858 at 1/1000 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.
Cytoplasm, nuclear membrane and nucleus staining on lymphocytes of Human tonsil tissue is observed.
Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody followed by ab97051 at 1/500.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling Bcl-2 with ab182858 at 1/250 (red) compared with a rabbit monoclonal IgG isotype control (ab172730) (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody (blue)). Goat anti rabbit IgG (FITC) at 1/500 was used as the secondary antibody.
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All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1 µg/ml
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 kDa
Observed band size: 26 kDaLanes 1 - 4: Merged signal (red and green). Green - ab182858 observed at 26 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab182858 was shown to specifically react with BCL2 when BCL2 knockout samples were used. Wild-type and BCL2 knockout samples were subjected to SDS-PAGE. Ab182858 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml 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.
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All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/10000 dilution
Lane 1 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 20 µg
Lane 2 : WEHI-3 (mouse leukemia cell line) whole cell lysate at 20 µg
Lane 3 : Mouse hippocampus at 10 µg
Lane 4 : Mouse heart at 10 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) at 1/2000 dilution
Predicted band size: 26 kDa
Observed band size: 26 kDa
Exposure time: 8 secondsBlocking/Diluting buffer 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] (ab182858)
Immunohistochemical analysis of paraffin-embedded Human endometrial cancer tissue labeling Bcl-2 with ab182858 at 1/1000 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.
Cytoplasm, nuclear membrane and nucleus staining on lymphocytes and cancer cells of Human endometrial cancer tissue is observed.
Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody followed by ab97051 at 1/500.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/20000 dilution
Lane 1 : Human tonsil lysate
Lane 2 : Human thymus lysate
Lane 3 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
Lane 4 : U-937 (Human histiocytic lymphoma cells) whole cell lysate
Lane 5 : THP-1 (Human monocytic leukemia cells) whole cell lysate
Lane 6 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 7 : C2C12 (Mouse myoblast cell line) whole cell lysate
Lane 8 : WEHI-3 (Mouse leukemia cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 26 kDa
Observed band size: 26 kDa
Exposure time: 1 minuteBlocking and diluting buffer was 5% NFDM /TBST.
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All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/2000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse kidney lysate
Lane 4 : Mouse spleen lysate
Lane 5 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 26 kDa
Observed band size: 26 kDa
Exposure time: 3 minutesBlocking and diluting buffer was 5% NFDM /TBST.
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All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/2000 dilution
Lane 1 : Human fetal kidney lysate
Lane 2 : Human fetal spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 26 kDa
Observed band size: 26 kDa
Exposure time: 3 minutesBlocking and diluting buffer was 5% NFDM /TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] (ab182858)
Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Bcl-2 with ab182858 at 1/1000 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.
Cytoplasm, nuclear membrane and nucleus staining on lymphocytes of Mouse spleen tissue is observed.
Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody followed by ab97051 at 1/500.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (458)
ab182858 被引用在 458 文献中.
- Wang L et al. Lactoferrin modification of berberine nanoliposomes enhances the neuroprotective effects in a mouse model of Alzheimer's disease. Neural Regen Res 18:226-232 (2023). PubMed: 35799547
- Ma J et al. Exosome-mediated lnc-ABCA12-3 promotes proliferation and glycolysis but inhibits apoptosis by regulating the toll-like receptor 4/nuclear factor kappa-B signaling pathway in esophageal squamous cell carcinoma. Korean J Physiol Pharmacol 27:61-73 (2023). PubMed: 36575934
- Wen Z et al. SIGIRR-caspase-8 signaling mediates endothelial apoptosis in Kawasaki disease. Ital J Pediatr 49:2 (2023). PubMed: 36600293
- Zhou M et al. PFKM inhibits doxorubicin-induced cardiotoxicity by enhancing oxidative phosphorylation and glycolysis. Sci Rep 12:11684 (2022). PubMed: 35804014
- Lu X et al. TSG-6 released from adipose stem cells-derived small extracellular vesicle protects against spinal cord ischemia reperfusion injury by inhibiting endoplasmic reticulum stress. Stem Cell Res Ther 13:291 (2022). PubMed: 35831906