Anti-Rad21抗体- ChIP Grade (ab992)

概述

  • 产品名称Anti-Rad21抗体- ChIP Grade
    参阅全部 Rad21 一抗
  • 描述
    兔多克隆抗体to Rad21 - ChIP Grade
  • 特异性The epitope recognized by ab992 maps to a region between residue 575 and the C-terminus (residue 631) human Rad21 homolog using the numbering given in entry NP_006256.1 (GeneID 5885).
  • 经测试应用适用于: IHC-P, ChIP, ICC/IF, WB, IPmore details
  • 种属反应性
    与反应: Mouse, Human, Xenopus laevis, Indian Muntjac
    预测可用于: Rat, Rabbit, Horse, Chicken, Guinea pig, Cow, Dog, Turkey, Chimpanzee, Gorilla, Chinese Hamster, Orangutan, Elephant
  • 免疫原

    Synthetic peptide (Human) conjugated to KLH - which represented a portion of human Rad21 encoded within exon 14 (LocusLink ID 5885).

  • 阳性对照
    • HeLa cell lysate. FFPE human breast fibroadenoma tissue sections.

性能

应用

Our Abpromise guarantee covers the use of ab992 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
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ChIP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

Used at a dilution of 1/200 for 30 min incubation (see Abreview for further information).

WB 1/500 - 1/5000. Predicted molecular weight: 72 kDa.

Band observed at ~130 kDa.

IP Use a concentration of 1 - 4 µg/ml.

靶标

  • 功能Cleavable component of the cohesin complex, involved in chromosome cohesion during cell cycle, in DNA repair, and in apoptosis. The cohesin complex is required for the cohesion of sister chromatids after DNA replication. The cohesin complex apparently forms a large proteinaceous ring within which sister chromatids can be trapped. At metaphase-anaphase transition, this protein is cleaved by separase/ESPL1 and dissociates from chromatin, allowing sister chromatids to segregate. The cohesin complex may also play a role in spindle pole assembly during mitosis. Also plays a role in apoptosis, via its cleavage by caspase-3/CASP3 or caspase-7/CASP7 during early steps of apoptosis: the C-terminal 64 kDa cleavage product may act as a nuclear signal to initiate cytoplasmic events involved in the apoptotic pathway.
  • 序列相似性Belongs to the rad21 family.
  • 结构域The C-terminal part associates with the head of SMC1A, while the N-terminal part binds to the head of SMC3.
  • 翻译后修饰Cleaved by separase/ESPL1 at the onset of anaphase. Cleaved by caspase-3 and caspase-7 at the beginning of apoptosis. The cleavage by ESPL1 and caspase-3 take place at different sites.
    Phosphorylated; becomes hyperphosphorylated in M phase of cell cycle. The large dissociation of cohesin from chromosome arms during prophase may be partly due to its phosphorylation by PLK.
  • 细胞定位Nucleus. Chromosome. Chromosome > centromere. Associates with chromatin. Before prophase it is scattered along chromosome arms. During prophase, most of cohesin complexes dissociate from chromatin probably because of phosphorylation by PLK, except at centromeres, where cohesin complexes remain. At anaphase, it is cleaved by separase/ESPL1, leading to the dissociation of the complex from chromosomes, allowing chromosome separation. Once cleaved by caspase-3, the C-terminal 64 kDa cleavage product translocates to the cytoplasm, where it may trigger apoptosis.
  • Information by UniProt
  • 数据库链接
  • 别名
    • CDLS4 antibody
    • Double-strand-break repair protein rad21 homolog antibody
    • hHR21 antibody
    • HR21 antibody
    • HRAD21 antibody
    • KIAA0078 antibody
    • MCD1 antibody
    • Nuclear matrix protein 1 antibody
    • NXP-1 antibody
    • NXP1 antibody
    • Protein involved in DNA double-strand break repair antibody
    • RAD21 antibody
    • RAD21 homolog (S. pombe) antibody
    • RAD21 homolog antibody
    • RAD21_HUMAN antibody
    • Scc1 antibody
    • SCC1 homolog antibody
    see all

Anti-Rad21 antibody - ChIP Grade 图像

  • Anti-Rad21 antibody - ChIP Grade (ab992) at 1/1000 dilution

    Predicted band size : 72 kDa
    Observed band size : 75 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 130 kDa (possible cross reactivity).
  • Anti-Rad21 antibody - ChIP Grade (ab992) at 1/2000 dilution + HeLa whole cell extract. Extraction was achieved using RIPA buffer and 1% SDS. 25 µg of total protein was loaded

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/1500 dilution (Anti-rabbit HRP (ab6721))

    Predicted band size : 72 kDa

    See Abreview

  • ab992 at a 1/200 dilution staining Asynchronous and paraformaldehyde-fixed (4%) HeLa cells by immunocytochemistry. The antibody was incubated with the cells 30 minutes and then detected using a Cy3 conjugated Goat Anti-Mouse IgG (H+L) antibody.

    This image is courtesy of an Abreview by Kirk McManus submitted on 27 February 2006.

    See Abreview

  • All lanes : Anti-Rad21 antibody - ChIP Grade (ab992) at 2 µg/ml

    Lane 1 : HeLa at 50 µg
    Lane 2 : HeLa at 15 µg
    Lane 3 : HeLa at 5 µg
    Lane 4 : 293T at 50 µg
    Lane 5 : NIH3T3 at 50 µg


    Predicted band size : 72 kDa


    Exposure time : 30 seconds
  • IHC image of Rad21 staining in human breast fibroadenoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab992, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Anti-Rad21 antibody - ChIP Grade (ab992)参考文献

This product has been referenced in:
  • Uusküla-Reimand L  et al. Topoisomerase II beta interacts with cohesin and CTCF at topological domain borders. Genome Biol 17:182 (2016). WB, ChIP . Read more (PubMed: 27582050) »
  • Majumder K  et al. Domain-Specific and Stage-Intrinsic Changes in Tcrb Conformation during Thymocyte Development. J Immunol 195:1262-72 (2015). ChIP ; Mouse . Read more (PubMed: 26101321) »

See all 38 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Blocking step Cells are incubated with primary antibody + BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 37°C
Sample Donkey Cell (Skin fibroblasts)
Specification Skin fibroblasts
Permeabilization Yes - 1% PFA + 0.2% Triton
Fixative Paraformaldehyde
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Abcam user community

Verified customer

提交于 Nov 12 2014

The epitope recognized by ab992 maps to a region between residue 575 and the C-terminus (residue 631) human Rad21 homolog using the numbering given in entry NP_006256.1 (GeneID 5885).

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Sample Mouse Cell (primary hepatoctyes)
Specification primary hepatoctyes
Permeabilization Yes - Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Fixative Paraformaldehyde
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Mr. Shengyun Ma

Verified customer

提交于 Oct 07 2013

Application Immunoprecipitation
Immuno-precipitation step Protein A
Sample Human Cell lysate - whole cell (Hep3B)
Specification Hep3B
Total protein in input 30 µg
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提交于 Sep 16 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Detection step Real-time PCR
Sample Mouse Cell lysate - whole cell (primary hepatoctyes)
Specification primary hepatoctyes
Negative control IgG
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldehyde
Positive control Suz12 on h19 ICR
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Abcam user community

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提交于 Sep 16 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 50 µg
Gel Running Conditions Non-reduced Denaturing (8%)
Sample Mouse Cell lysate - whole cell (Primary hepatoctyes)
Specification Primary hepatoctyes
Blocking step I-Block(Applied biosystems) as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 2µg/mL · Temperature: 4°C
Username

Mr. Shengyun Ma

Verified customer

提交于 Aug 09 2013



Based on Swiss-prot, the predicted molecular weight for this protein is indeed 75 kDa as mentioned on our datasheet. However, a band at ˜120-130 kDa has been previously observed: Mol Cell Biol. 2002 Dec;22(23):8267-77 PMID: 12417729. The high...

Read More

I've compiled some information below regarding potential primer sets for the ChIP antibodies we talked about, where information was lacking (verified in ChIP by other researches), I included links to abReviews and references that used the antibody in C...

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application CHIPseq
Sample Human Cell lysate - nuclear (Raji)
Specification Raji
Username

Abcam user community

Verified customer

提交于 Jan 07 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - nuclear (Raji)
Specification Raji
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 7 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1 % Formaldehyd
Detection step Real-time PCR
Positive control EBV specific locus(EBV 3+)
Negative control EBV specific locus, GAPDH, Centromeric region of chromosome 1 (Cen)
Username

Abcam user community

Verified customer

提交于 Jan 07 2013

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