Anti-CLLD8/SETDB2抗体(ab5517)
Key features and details
- Rabbit polyclonal to CLLD8/SETDB2
- Suitable for: WB, ICC/IF
- Reacts with: Human
- Isotype: IgG
概述
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产品名称
Anti-CLLD8/SETDB2抗体
参阅全部 CLLD8/SETDB2 一抗 -
描述
兔多克隆抗体to CLLD8/SETDB2 -
宿主
Rabbit -
经测试应用
适用于: WB, ICC/IFmore details -
种属反应性
与反应: Human -
免疫原
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常规说明
Our collaborator on this project has found that the antibody works poorly in WB.Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
存储溶液
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
纯度
Immunogen affinity purified -
克隆
多克隆 -
同种型
IgG -
研究领域
相关产品
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Compatible Secondaries
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Isotype control
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Recombinant Protein
应用
Our Abpromise guarantee covers the use of ab5517 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB | Use at an assay dependent concentration. Predicted molecular weight: 82 kDa. | |
| ICC/IF | Use at an assay dependent concentration. |
靶标
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功能
Histone methyltransferase involved in left-right axis specification in early development and mitosis. Specifically trimethylates 'Lys-9' of histone H3 (H3K9me3). H3K9me3 is a specific tag for epigenetic transcriptional repression that recruits HP1 (CBX1, CBX3 and/or CBX5) proteins to methylated histones. Contributes to H3K9me3 in both the interspersed repetitive elements and centromere-associated repeats. Plays a role in chromosome condensation and segregation during mitosis. -
组织特异性
Ubiquitous. Highest expression in heart, testis and ovary. -
序列相似性
Belongs to the histone-lysine methyltransferase family.
Contains 1 MBD (methyl-CpG-binding) domain.
Contains 1 pre-SET domain.
Contains 1 SET domain. -
细胞定位
Nucleus. Chromosome. - Information by UniProt
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数据库链接
- Entrez Gene: 83852 Human
- Omim: 607865 Human
- SwissProt: Q96T68 Human
- Unigene: 631789 Human
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别名
- C13orf4 antibody
- Chromosome 13 open reading frame 4 antibody
- Chronic lymphocytic leukemia deletion region 8 antibody
see all
图片
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Western blot - Anti-CLLD8/SETDB2 antibody (ab5517)Anti-CLLD8/SETDB2 antibody (ab5517) at 1 µg/ml +Recombinant Human CLLD8/SETDB2 protein (ab169554) at 0.01 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 82 kDa
Additional bands at: 120 kDa (possible tagged protein)
Exposure time: 10 secondsab5517 recognises a band corresponding to the tagged CLLD8/SETB2 at approximately 120 kDa.
Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
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Immunocytochemistry/ Immunofluorescence - Anti-CLLD8/SETDB2 antibody (ab5517)ICC/IF image of ab5517 stained MCF-7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab5517 at 5ug/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 Goat anti Rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Immunocytochemistry/ Immunofluorescence - Anti-CLLD8/SETDB2 antibody (ab5517)This image is courtesy of Genevieve Fourel, GrenobleLeft: Endogenous CLLD8
Detection using indirect fluorescence of the signal corresponding to endogenous CLLD8 in 293T cells. Cells fixed using 4% formaldehyde, blocked with PBS containing 3% milk and 0.5% Triton X-100, incubated for 1 hour at 37 °C using a 1/50 dilution of antibody ab5517. Cells were then washed 3 times and incubated for 1 hour at 37 °C with a goat anti-rabbit secondary antibody (1/500 dilution) coupled to Alexa Fluor 555 (Molecular Probes). Following 3 more washes, cells were stained with DAPI and mounted with Vectashield.
Top: DAPI
Bottom: ab5517
Right: Overexpressed CLLD8Detection using indirect fluorescence of the signal corresponding to staining with anti-FLAG mouse antibody (top) and an antibody (ab5517) against CLLD8 (middle) in 293T cells. 293T cells were transfected with vectors for overexpression of flagged CLLD8 using Lipofectamine 2000 transfection procedure (Invitrogen). Cells were fixed 48 hours post-tra
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Immunocytochemistry/ Immunofluorescence - Anti-CLLD8/SETDB2 antibody (ab5517)This image is courtesy of Genevieve Fourel, GrenobleDetection using indirect fluorescence of the signal corresponding to endogenous CLLD8 in 293T cells. Using Lipofectamine 2000 (Invitrogen), cells were transfected with either a control shRNA directed against luciferase (left, SiRluc) or a specific siRNA directed against CLLD8 (right). 48 hours post-transfection, cells were fixed, blocked, and incubated with a 1:50 dilution of ab5517 at 37 °C for 1 hour. Cells were then washed 3 times and incubated at 37 °C for 1 hour with a 1/500 dilution of goat anti-rabbit antibody coupled with Alexa Fluor 555 (Molecular Probes). Following 3 further washes cells were stained with DAPI and mounted with Vectashield.
Antibody signals were extinguished when a specific RNAi was used. The few cells which retained a signal were presumably not transfected.
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Western blot - Anti-CLLD8/SETDB2 antibody (ab5517)All lanes : Anti-CLLD8/SETDB2 antibody (ab5517) at 1 µg/ml
Lane 1 : Sf21 cells infected by baculovirus containing FLAG-CLLD8 gene at 10 µg
Lane 2 : Sf21 cells infected by baculovirus containing FLAG-CLLD8 gene at 20 µg
Lane 3 : non-infected Sf21 cells at 10 µg
Lane 4 : non-infected Sf21 cells at 20 µg
Lane 5 : Sf21 infected by baculovirus containing FLAG-CLLD8 gene at 10 µg with Human CLLD8/SETDB2 peptide (ab24398) at 1 µg/ml
Lane 6 : Sf21 infected by baculovirus containing FLAG-CLLD8 gene at 20 µg with Human CLLD8/SETDB2 peptide (ab24398) at 1 µg/ml
Lane 7 : non-infected Sf21 cells at 10 µg with Human CLLD8/SETDB2 peptide (ab24398) at 1 µg/ml
Lane 8 : non-infected Sf21 cells at 20 µg with Human CLLD8/SETDB2 peptide (ab24398) at 1 µg/ml
Predicted band size: 82 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?
Additional bands at: 22 kDa (possible non-specific binding), 90 kDa (possible non-specific binding)
ab5517 recognises a band corresponding to FLAG-tagged CLLD8/SETB2 at approximately 110 kDa in Sf21 cells infected by baculovirus expressing CLLD8/SETDB2 (lanes1-2). This is larger than the predicated band size (81 kDa) due to the addition of the FLAG tag.
ab5517 does not detect a band in uninfected sf21 cells (lanes3-4) or following blocking studies using the immunizing peptide (lanes5-6).
There appears to be a ladder of proteins recognised by ab5517 (lanes1-2), which could be attributed to degradation products.
实验方案
数据表及文件
文献 (3)
ab5517 被引用在 3 文献中.
- Nishikawaji T et al. Oncogenic roles of the SETDB2 histone methyltransferase in gastric cancer. Oncotarget 7:67251-67265 (2016). IHC . PubMed: 27572307
- Hogarth CA et al. Identification and expression of potential regulators of the mammalian mitotic-to-meiotic transition. Biol Reprod 84:34-42 (2011). WB, IHC-P ; Mouse . PubMed: 20826732
- Falandry C et al. CLLD8/KMT1F is a lysine methyltransferase that is important for chromosome segregation. J Biol Chem 285:20234-41 (2010). WB, ICC/IF ; Human . PubMed: 20404330
