重组Anti-MCL1抗体[Y37] (ab32087)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y37] to MCL1
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-MCL1抗体[Y37]
参阅全部 MCL1 一抗 -
描述
兔单克隆抗体[Y37] to MCL1 -
宿主
Rabbit -
特异性
ab32087 recognises MCL1. The antibody does not cross-react with other Bcl-2 family members. -
经测试应用
适用于: Flow Cyt (Intra), ICC/IF, WB, IHC-P, IPmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
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阳性对照
- WB: Human lung, lung cancer and liver lysates; HEK293T, A431, Ramos, H:-60, HeLa, MCF7 and HepG2 whole cell lysate (ab7900). IHC-P: Human colon adenocarcinoma tissue. ICC/IF: HCT 116, MCF7 and H1299 cells. Flow Cyt (intra): Ramos and A431 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.
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.
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. -
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
Y37 -
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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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32087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.
For unpurified use 1 ug for 106 cells. (For lot-specific stock concentration, please contact Abcam).ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF | (2) |
1/100 - 1/500.
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WB | (10) |
1/1000 - 1/5000. Predicted molecular weight: 37 kDa.
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IHC-P |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP | (1) |
Use at an assay dependent concentration.
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说明 |
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Flow Cyt (Intra)
1/250. For unpurified use 1 ug for 106 cells. (For lot-specific stock concentration, please contact Abcam).ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/100 - 1/500. |
WB
1/1000 - 1/5000. Predicted molecular weight: 37 kDa. |
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
靶标
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功能
Involved in the regulation of apoptosis versus cell survival, and in the maintenance of viability but not of proliferation. Mediates its effects by interactions with a number of other regulators of apoptosis. Isoform 1 inhibits apoptosis. Isoform 2 promotes apoptosis. -
序列相似性
Belongs to the Bcl-2 family. -
翻译后修饰
Cleaved by CASP3 during apoptosis. In intact cells cleavage occurs preferentially after Asp-127, yielding a pro-apoptotic 28 kDa C-terminal fragment.
Rapidly degraded in the absence of phosphorylation on Thr-163 in the PEST region.
Phosphorylated on Thr-163. Treatment with taxol or okadaic acid induces phosphorylation on additional sites. -
细胞定位
Membrane. Cytoplasm. Mitochondrion. Nucleus > nucleoplasm. Cytoplasmic, associated with mitochondria. - Information by UniProt
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数据库链接
- Entrez Gene: 4170 Human
- Entrez Gene: 17210 Mouse
- Entrez Gene: 60430 Rat
- Omim: 159552 Human
- SwissProt: Q07820 Human
- SwissProt: P97287 Mouse
- SwissProt: Q9Z1P3 Rat
- Unigene: 632486 Human
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别名
- Bcl 2 related protein EAT/mcl1 antibody
- Bcl-2-like protein 3 antibody
- Bcl-2-related protein EAT/mcl1 antibody
see all
图片
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All lanes : Anti-MCL1 antibody [Y37] (ab32087) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : MCL1 knockout HEK293T cell lysate
Lane 3 : Ramos cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 37 kDa
Observed band size: 37 kDaLanes 1-3: Merged signal (red and green). Green - ab32087 observed at 37 kDa. Red - loading control ab7291 observed at 50 kDa.
ab32087 Anti-MCL1 antibody [Y37] was shown to specifically react with MCL1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266838 (knockout cell lysate ab256986) was used. Wild-type and MCL1 knockout samples were subjected to SDS-PAGE. ab32087 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 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-MCL1 antibody [Y37] (ab32087)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon adenocarcinoma tissue labelling MCL1 with purified ab32087 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunocytochemistry/Immunofluorescence analysis of HCT 116 (human colorectal carcinoma cell line) cells labelling MCL1 with purified ab32087 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).
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Intracellular Flow Cytometry analysis ofRamos (human Burkitt's lymphoma cell line) cells labelling MCL1 with purified ab32087 at 1/250 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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All lanes : Anti-MCL1 antibody [Y37] (ab32087) at 1/1000 dilution
Lane 1 : Human lung tissue with NFDM/TBST
Lane 2 : Human lung cancer tissue with NFDM/TBST
Lane 3 : Human liver tissue with NFDM/TBST
Lysates/proteins at 20 µg per lane.
Blocking peptides at 5 % per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (H+L), Peroxidase conjugated. at 1/1000 dilution
Predicted band size: 37 kDaExposure time for samples 1-3: 15 seconds; exposure time for samples 4-6: 1 minute.
Additional bands: We are unsure as to the identity of these extra bands.
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All lanes : Anti-MCL1 antibody [Y37] (ab32087) at 1/1000 dilution
Lane 1 : Ramos (human Burkitt's lymphoma cell line) cell lysate with NFDM/TBST
Lane 2 : HL-60 (human promyelocytic leukemia cell line) cell lysate with NFDM/TBST
Lane 3 : A431 (human epidermoid carcinoma cell line) cell lysate with NFDM/TBST
Lane 4 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate with NFDM/TBST
Lane 5 : MCF7 (human breast adenocarcinoma cell line) cell lysate with NFDM/TBST
Lane 6 : HepG2 (human liver hepatocellular carcinoma cell line) cell lysate with NFDM/TBST
Lysates/proteins at 20 µg per lane.
Blocking peptides at 5 % per lane.
Secondary
All lanes : Goat anti-rabbit IgG (H+L), peroxidase conjugated. at 1/1000 dilution
Predicted band size: 37 kDa
Exposure time: 15 secondsAdditional bands: We are unsure as to the identity of these extra bands.
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Immunocytochemistry/ Immunofluorescence - Anti-MCL1 antibody [Y37] (ab32087)This image is courtesy of an anonymous Abreview.
Immunocytochemistry/Immunofluorescence analysis of H1299 cells labelling MCL1 with unpurified ab32087. Cells were PFA-fixed and permeabilized in 0.5% Triton X-100 prior to blocking in 3% Serum for 1 hour at 24°C. The primary antibody was diluted 1/100 and incubated with the sample for 1 hour at 24°C. The secondary antibody was an Alexa Fluor® 488-conjugated Goat anti-Rabbit polyclonal, diluted 1/2000. DAPI (blue) was used as the nuclear counterstain.
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Intracellular Flow Cytometry analysis ofA431 (human epidermoid carcinoma cell line) cells labelling MCL1 with unpurified ab32087 (red line). Cells were fixed with methanol (5 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 (ab32087, 1µg/1x106 cells) 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.
Black - Isotype control, rabbit monoclonal IgG.
Acquisition of >5,000 events was performed. This antibody gave a decreased signal in A431 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.
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Immunocytochemistry/Immunofluorescence analysis of HCT 116 (human colorectal carcinoma cell line) cells treated with wogonin (ab142471) labelling MCL1 with unpurified ab32087. Decrease of MCL1 expression correlates with increased concentration of wogonin, as described in literature. Cells were incubated at 37°C for 2h in media containing different concentrations of ab142471 (wogonin) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab32087 (1/100) dilution was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
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 (138)
ab32087 被引用在 138 文献中.
- Song Y et al. β-Hydroxybutyrate inhibits apoptosis in bovine neutrophils through activating ERK1/2 and AKT signaling pathways. J Dairy Sci 105:3477-3489 (2022). PubMed: 35151471
- He L et al. CDK7/12/13 inhibition targets an oscillating leukemia stem cell network and synergizes with venetoclax in acute myeloid leukemia. EMBO Mol Med 14:e14990 (2022). PubMed: 35253392
- Koessinger AL et al. Increased apoptotic sensitivity of glioblastoma enables therapeutic targeting by BH3-mimetics. Cell Death Differ 29:2089-2104 (2022). PubMed: 35473984
- Zhu X et al. Ordered micropattern arrays fabricated by lung-derived dECM hydrogels for chemotherapeutic drug screening. Mater Today Bio 15:100274 (2022). PubMed: 35601895
- Haiyilati A et al. Gga-miR-30c-5p Enhances Apoptosis in Fowl Adenovirus Serotype 4-Infected Leghorn Male Hepatocellular Cells and Facilitates Viral Replication through Myeloid Cell Leukemia-1. Viruses 14:N/A (2022). PubMed: 35632731