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: ICC/IF, WB, IHC-P, Flow Cyt
- Knockout validated
- Reacts with: Human
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. -
Tested Applications & Species
Application Species Flow Cyt HumanICC/IF HumanIHC-P HumanWB 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: 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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
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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KO cell pellets
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Positive Controls
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32087 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
应用 | Species |
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Flow Cyt |
Human
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ICC/IF |
Human
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IHC-P |
Human
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WB |
Human
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All applications |
Mouse
Rat
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应用 | Ab评论 | 说明 |
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ICC/IF | (2) |
1/100 - 1/500.
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WB | (6) |
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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Flow Cyt |
1/250.
For unpurified use 1 ug for 106 cells. (For lot-specific stock concentration, please contact Abcam). |
说明 |
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ICC/IF
1/100 - 1/500. |
WB
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. |
Flow Cyt
1/250. For unpurified use 1 ug for 106 cells. (For lot-specific stock concentration, please contact Abcam). |
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
see all -
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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Flow Cytometry analysis of Ramos (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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Flow Cytometry analysis of A431 (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 (87)
ab32087 被引用在 87 文献中.
- Knoll G et al. NOXA-dependent contextual synthetic lethality of BCL-XL inhibition and "osmotic reprogramming" in colorectal cancer. Cell Death Dis 11:257 (2020). PubMed: 32312973
- Yuan B et al. Role of Bcl-2 on drug resistance in breast cancer polyploidy-induced spindle poisons. Oncol Lett 19:1701-1710 (2020). PubMed: 32194662
- Xia P et al. MicroRNA-377 exerts a potent suppressive role in osteosarcoma through the involvement of the histone acetyltransferase 1-mediated Wnt axis. J Cell Physiol 234:22787-22798 (2019). PubMed: 31152456
- Du Q et al. Propofol inhibits proliferation and epithelial-mesenchymal transition of MCF-7 cells by suppressing miR-21 expression. Artif Cells Nanomed Biotechnol 47:1265-1271 (2019). PubMed: 30942630
- Liu B et al. LncRNA ANRIL protects against oxygen and glucose deprivation (OGD)-induced injury in PC-12 cells: potential role in ischaemic stroke. Artif Cells Nanomed Biotechnol 47:1384-1395 (2019). PubMed: 31174432