Anti-Rad21抗体(ab992)

Reviews (19)Q&A (9)References (134)
Western blot - Anti-Rad21 antibody (ab992)
  • Western blot - Anti-Rad21 antibody (ab992)
  • Western blot - Anti-Rad21 antibody (ab992)

Key features and details

  • Rabbit polyclonal to Rad21
  • Suitable for: WB
  • Reacts with: Mouse, Human
  • Isotype: IgG

概述

  • 产品名称

    Anti-Rad21抗体
    参阅全部 Rad21 一抗

  • 描述

    兔多克隆抗体to Rad21

  • 宿主

    Rabbit

  • 特异性

    The epitope recognized by Anti-Rad21 antibody (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).

  • Tested Applications & Species

    Application Species
    WB
    Human
    See all applications and species data

  • 免疫原

    Synthetic peptide corresponding to Human Rad21 (C terminal) conjugated to keyhole limpet haemocyanin. Represented a portion of human Rad21 encoded within exon 14 (LocusLink ID 5885).

  • 常规说明

      

    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

应用

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

应用 Species
WB
Human
All applications
Rat
Rabbit
Horse
Chicken
Guinea pig
Cow
Dog
Turkey
Chimpanzee
Gorilla
Chinese hamster
Orangutan
Elephant
应用 Ab评论 说明
WB (7)
1/500 - 1/5000. Predicted molecular weight: 72 kDa.

Band observed at ~130 kDa.
说明
WB
1/500 - 1/5000. Predicted molecular weight: 72 kDa.

Band observed at ~130 kDa.

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

    see all

  • 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

图片

  • Western blot - Anti-Rad21 antibody (ab992)
    Western blot - Anti-Rad21 antibody (ab992)
    Anti-Rad21 antibody (ab992) at 1/1000 dilution + HeLa whole cell lysate

    Developed using the ECL technique.

    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)

  • Western blot - Anti-Rad21 antibody (ab992)
    Western blot - Anti-Rad21 antibody (ab992)
    Anti-Rad21 antibody (ab992) at 1/2000 dilution + HeLa whole cell extract (extraction was achieved using RIPA buffer and 1% SDS) at 25 µg/ml

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

    Predicted band size: 72 kDa



    A rad21 knock down negative control was employed.

    This image is courtesy of an Abreview submitted on 9 September 2005. We do not have any further information relating to this image.

    See Abreview

  • Western blot - Anti-Rad21 antibody (ab992)
    Western blot - Anti-Rad21 antibody (ab992)
    All lanes : Anti-Rad21 antibody (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

文献 (134)

发表研究结果有使用 ab992？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab992 被引用在 134 文献中.

  • Kentepozidou E  et al. Clustered CTCF binding is an evolutionary mechanism to maintain topologically associating domains. Genome Biol 21:5 (2020). PubMed: 31910870
  • Khoury A  et al. Constitutively bound CTCF sites maintain 3D chromatin architecture and long-range epigenetically regulated domains. Nat Commun 11:54 (2020). PubMed: 31911579
  • Jia Z  et al. Tandem CTCF sites function as insulators to balance spatial chromatin contacts and topological enhancer-promoter selection. Genome Biol 21:75 (2020). PubMed: 32293525
  • McNicoll F  et al. Meiotic sex chromosome cohesion and autosomal synapsis are supported by Esco2. Life Sci Alliance 3:N/A (2020). PubMed: 32051254
  • Nishana M  et al. Defining the relative and combined contribution of CTCF and CTCFL to genomic regulation. Genome Biol 21:108 (2020). PubMed: 32393311
View all Publications for this product

客户评价及客户问答

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1-10 of 28 Abreviews or Q&A

Question

Customer kindly contacted us regarding positve and negative primers for our ChIP grade antibodies.

Read More
Answer

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 ChIP.

1. ab817: GAPDH (+), GAD1 (-), AFM (-), Factor VIII (-)

2. ab1791: GAPDH, RPL30, ALDOA, MYO-D, SERPINA, AFM, SAT2, SATa (each batch is verified against this primer set, an example image of one batch test is given on the datasheet)

3. ab7030: was verified in ChIP by other researchers submitting data through our abReview system and specific references. Their positive and negative controls used are given through the link below, as well as a "click to contact this user" link that allows you to contact the researchers if you would like more specific primer information. The specific references with ChIP listed in bold are good sources for positive and negative primers as well.
a) Abreview: (NFKBIA promoter (+), GAPDH (-)) https://www.abcam.com/HDAC3-antibody-ChIP-Grade-ab7030-reviews.html?intabreviewid=25057
b) Abreview: https://www.abcam.com/HDAC3-antibody-ChIP-Grade-ab7030-reviews.html?intabreviewid=18386
c) Specific references: https://www.abcam.com/HDAC3-antibody-ChIP-Grade-ab7030-references.html

4. ab111069 Untested in ChIP.

5. ab992: was verified in ChIP by other researchers (abReviews and specific references)
a) Specific references: https://www.abcam.com/Rad21-antibody-ChIP-Grade-ab992-references.html
b) Abreview: https://www.abcam.com/Rad21-antibody-ChIP-Grade-ab992-reviews.html?intabreviewid=14542
c) Abreview: (EBV 3+ (+), EBV1,2,4,5,6 (-), GAPDH (-), Cen (-)) https://www.abcam.com/Rad21-antibody-ChIP-Grade-ab992-reviews.html?intabreviewid=33942

Our ChIP primer sets:
https://www.abcam.com/index.html?pageconfig=resource&rid=10600

Read More

ChIP abreview for Anti-Rad21 antibody - ChIP Grade

 Good
Abreviews
Application
ChIP
Sample
Human Cell lysate - nuclear (MCF10a)
Negative control
Random regions without ATAC-seq signal or any histone marks.
Specification
MCF10a
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Positive control
Strong CTCF binding sites
Read More

Abcam user community

Verified customer

提交于 Dec 10 2019

ChIP abreview for Anti-Rad21 antibody - ChIP Grade

 Below Average
Abreviews
abreview image
Application
ChIP
Sample
Mouse Tissue lysate - nuclear (Limb buds, forebrain, ESC)
Negative control
We perform ChIP-seq
Specification
Limb buds, forebrain, ESC
Detection step
Other
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 37% Formaldehyde (Sigma), dilu
Positive control
We perform ChIP-seq
Read More

Daniel Ibrahim

Verified customer

提交于 Nov 12 2018

Western blot abreview for Anti-Rad21 antibody - ChIP Grade

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Whole cell lysate resolved on a 12% SDS PAGE gel)
Gel Running Conditions
Reduced Denaturing (12% SDS gel)
Loading amount
25 µg
Treatment
HEK293T cells transfected with HA-Rad21 plasmid for overexpression
Specification
Whole cell lysate resolved on a 12% SDS PAGE gel
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

提交于 Jan 19 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-Rad21 antibody - ChIP Grade

 Good
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

提交于 Nov 12 2014

Question

What does this antibody recognize on human Rad21?

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Answer

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).

Read More

Immunocytochemistry/ Immunofluorescence abreview for Anti-Rad21 antibody - ChIP Grade

 Good
Abreviews
abreview image
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
Read More

Mr. Shengyun Ma

Verified customer

提交于 Oct 07 2013

Immunoprecipitation abreview for Anti-Rad21 antibody - ChIP Grade

 Below Average
Abreviews
abreview image
Application
Immunoprecipitation
Immuno-precipitation step
Protein A
Sample
Human Cell lysate - whole cell (Hep3B)
Specification
Hep3B
Total protein in input
30 µg
Read More

Abcam user community

Verified customer

提交于 Sep 16 2013

ChIP abreview for Anti-Rad21 antibody - ChIP Grade

 Good
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

提交于 Sep 16 2013

Western blot abreview for Anti-Rad21 antibody - ChIP Grade

 Average
Abreviews
abreview image
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
Read More

Mr. Shengyun Ma

Verified customer

提交于 Aug 09 2013

1-10 of 28 Abreviews or Q&A

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