製品の概要
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製品名Anti-Rad21 antibody - ChIP Grade
Rad21 一次抗体 製品一覧 -
製品の詳細Rabbit polyclonal to Rad21 - ChIP Grade
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由来種Rabbit
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特異性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).
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アプリケーション適用あり: IHC-P, ChIP, ICC/IF, WB, IPmore details
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種交差性交差種: Mouse, Human, Xenopus laevis, Indian muntjac
交差が予測される動物種: Rat, Rabbit, Horse, Chicken, Guinea pig, Cow, Dog, Turkey, Chimpanzee, Gorilla, Chinese hamster, Orangutan, Elephant
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免疫原
Synthetic peptide (Human) conjugated to KLH - which represented a portion of human Rad21 encoded within exon 14 (LocusLink ID 5885).
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ポジティブ･コントロール
- HeLa cell lysate. FFPE human breast fibroadenoma tissue sections.
製品の特性
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製品の状態Liquid
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保存方法Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
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バッファーPreservative: 0.1% Sodium azide
Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris -
Concentration information loading... -
精製度Immunogen affinity purified
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ポリ/モノポリクローナル
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アイソタイプIgG
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研究分野
関連製品
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ChIP Related Products
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Compatible Secondaries
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Immunohistochemistry kits
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Related Products
アプリケーション
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.
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| 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). |
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| WB | 1/500 - 1/5000. Predicted molecular weight: 72 kDa. Band observed at ~130 kDa. |
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| IP | Use a concentration of 1 - 4 µg/ml. |
ターゲット情報
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機能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.
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配列類似性Belongs to the rad21 family.
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ドメインThe C-terminal part associates with the head of SMC1A, while the N-terminal part binds to the head of SMC3.
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翻訳後修飾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. -
細胞内局在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
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参照データベース
- Entrez Gene: 540966 Cow
- Entrez Gene: 5885 Human
- Entrez Gene: 19357 Mouse
- Entrez Gene: 314949 Rat
- Entrez Gene: 399129 Xenopus laevis
- Omim: 606462 Human
- SwissProt: Q3SWX9 Cow
- SwissProt: O60216 Human
see all -
別名
- CDLS4 antibody
- Double-strand-break repair protein rad21 homolog antibody
- hHR21 antibody
see all
画像
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Western blot - Anti-Rad21 antibody - ChIP Grade (ab992)Anti-Rad21 antibody - ChIP Grade (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) -
Immunocytochemistry/ Immunofluorescence - Anti-Rad21 antibody - ChIP Grade (ab992)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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad21 antibody - ChIP Grade (ab992)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. -
Western blot - Anti-Rad21 antibody - ChIP Grade (ab992)Anti-Rad21 antibody - ChIP Grade (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. -
Western blot - Anti-Rad21 antibody - ChIP Grade (ab992)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
プロトコール
参考文献
This product has been referenced in:
- Zheng G et al. MCM2-7-dependent cohesin loading during S phase promotes sister-chromatid cohesion. Elife 7:N/A (2018). Read more (PubMed: 29611806) »
- Matthews BJ & Waxman DJ Computational prediction of CTCF/cohesin-based intra-TAD loops that insulate chromatin contacts and gene expression in mouse liver. Elife 7:N/A (2018). Read more (PubMed: 29757144) »
レビューと Q&A
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