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  1. Link

    kap1-antibody-ab10483.pdf

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Epigenetics and Nuclear Signaling Chromatin Binding Proteins Methylated DNA
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Anti-KAP1 抗体 (ab10483)

  • Datasheet
  • SDS
Reviews (7)Q&A (2)References (59)

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Abpromise

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAP1 antibody (ab10483)
  • Immunoprecipitation - Anti-KAP1 antibody (ab10483)
  • Western blot - Anti-KAP1 antibody (ab10483)
  • Western blot - Anti-KAP1 antibody (ab10483)
  • Immunoprecipitation - Anti-KAP1 antibody (ab10483)

Key features and details

  • Rabbit polyclonal to KAP1
  • Suitable for: WB, IHC-P, IP
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-KAP1 antibody [EPR5249] (ab109545)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 製品名

    Anti-KAP1 antibody
    KAP1 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to KAP1
  • 由来種

    Rabbit
  • アプリケーション

    適用あり: WB, IHC-P, IPmore details
  • 種交差性

    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide. The immunogen is between aa 1-50.
    Database link: Q13263

  • ポジティブ・コントロール

    • ウェスタン・ブロット用ポジティブコントロールRecombinant Human KAP1 protein
    • WB: human HeLa, HEK293T cells; mouse NIH3T3, TCMK-1, 4T1, CT26.WT, rat C6 cells. IP: HeLa cells. IHC: human breast carcinoma
  • 特記事項

    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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

製品の特性

  • 製品の状態

    Liquid
  • 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.
  • バッファー

    pH: 7
    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
  • 特記事項(精製)

    Antibodies were affinity purified using the peptide immobilized on solid support.
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • Epigenetics and Nuclear Signaling
    • Chromatin Binding Proteins
    • Methylated DNA
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ubiquitin E3 Enzymes
    • RING Finger E3 Ligase
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Other
    • Cancer
    • Cell Death
    • Apoptosis
    • Other
    • Epigenetics and Nuclear Signaling
    • Bromodomains

関連製品

  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • HEK-293 whole cell lysate (ab7902)
  • Recombinant Protein

    • Recombinant Human KAP1 protein (ab131899)

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab10483の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB (3)
1/2000 - 1/10000. Detects a band of approximately 110 kDa (predicted molecular weight: 100 kDa).
IHC-P (1)
1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP (1)
Use at 2-10 µg/mg of lysate.
特記事項
WB
1/2000 - 1/10000. Detects a band of approximately 110 kDa (predicted molecular weight: 100 kDa).
IHC-P
1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
Use at 2-10 µg/mg of lysate.

ターゲット情報

  • 機能

    Nuclear corepressor for KRAB domain-containing zinc finger proteins (KRAB-ZFPs). Mediates gene silencing by recruiting CHD3, a subunit of the nucleosome remodeling and deacetylation (NuRD) complex, and SETDB1 (which specifically methylates histone H3 at 'Lys-9' (H3K9me)) to the promoter regions of KRAB target genes. Enhances transcriptional repression by coordinating the increase in H3K9me, the decrease in histone H3 'Lys-9 and 'Lys-14' acetylation (H3K9ac and H3K14ac, respectively) and the disposition of HP1 proteins to silence gene expression. Recruitment of SETDB1 induces heterochromatinization. May play a role as a coactivator for CEBPB and NR3C1 in the transcriptional activation of ORM1. Also corepressor for ERBB4. Inhibits E2F1 activity by stimulating E2F1-HDAC1 complex formation and inhibiting E2F1 acetylation. May serve as a partial backup to prevent E2F1-mediated apoptosis in the absence of RB1. Important regulator of CDKN1A/p21(CIP1). Has E3 SUMO-protein ligase activity toward itself via its PHD-type zinc finger.
  • 組織特異性

    Expressed in all tissues tested including spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes.
  • パスウェイ

    Protein modification; protein sumoylation.
  • 配列類似性

    Belongs to the TRIM/RBCC family.
    Contains 2 B box-type zinc fingers.
    Contains 1 bromo domain.
    Contains 1 PHD-type zinc finger.
    Contains 1 RING-type zinc finger.
  • ドメイン

    The HP1 box is both necessary and sufficient for HP1 binding.
    The PHD-type zinc finger enhances CEBPB transcriptional activity. The PHD-type zinc finger, the HP1 box and the bromo domain, function together to assemble the machinery required for repression of KRAB domain-containing proteins. Acts as an intramolecular SUMO E3 ligase for autosumoylation of bromodomain.
    The RING-finger-B Box-coiled-coil/tripartite motif (RBCC/TRIM motif) is required for interaction with the KRAB domain of KRAB-zinc finger proteins. Binds four zinc ions per molecule. The RING finger and the N-terminal of the leucine zipper alpha helical coiled-coil region of RBCC are required for oligomerization.
    Contains one Pro-Xaa-Val-Xaa-Leu (PxVxL) motif, which is required for interaction with chromoshadow domains. This motif requires additional residues -7, -6, +4 and +5 of the central Val which contact the chromoshadow domain.
  • 翻訳後修飾

    Phosphorylated upon DNA damage, probably by ATM or ATR. ATM-induced phosphorylation on Ser-824 represses sumoylation leading to the de-repression of expression of a subset of genes involved in cell cycle control and apoptosis in response to genotoxic stress. Dephosphorylation by the phosphatases, PPP1CA and PP1CB forms, allows sumoylation and expression of TRIM28 target genes.
    Sumoylation/desumoylation events regulate TRIM28-mediated transcriptional repression. Sumoylation is required for interaction with CHD3 and SETDB1 and the corepressor activity. Represses and is repressed by Ser-824 phosphorylation. Enhances the TRIM28 corepressor activity, inhibiting transcriptional activity of a number of genes including GADD45A and CDKN1A/p21. Lys-554, Lys-779 and Lys-804 are the major sites of sumoylation. In response to Dox-induced DNA damage, enhanced phosphorylation on Ser-824 prevents sumoylation and allows de-repression of CDKN1A/p21.
  • 細胞内局在

    Nucleus. Associated with centromeric heterochromatin during cell differentiation through CBX1.
  • Target information above from: UniProt accession Q13263 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 参照データベース

    • Entrez Gene: 10155 Human
    • Entrez Gene: 21849 Mouse
    • Entrez Gene: 116698 Rat
    • Omim: 601742 Human
    • SwissProt: Q13263 Human
    • SwissProt: Q62318 Mouse
    • SwissProt: O08629 Rat
    • Unigene: 467408 Human
    • Unigene: 15701 Mouse
    • Unigene: 398345 Mouse
    • Unigene: 198494 Rat
    see all
  • 別名

    • E3 SUMO protein ligase TRIM28 antibody
    • E3 SUMO-protein ligase TRIM28 antibody
    • FLJ29029 antibody
    • KAP 1 antibody
    • KAP-1 antibody
    • KRAB associated protein 1 antibody
    • KRAB interacting protein 1 antibody
    • KRAB-associated protein 1 antibody
    • KRAB-interacting protein 1 antibody
    • KRIP 1 antibody
    • KRIP-1 antibody
    • KRIP1 antibody
    • Nuclear corepressor KAP 1 antibody
    • Nuclear corepressor KAP-1 antibody
    • RING finger protein 96 antibody
    • RNF96 antibody
    • TF1B antibody
    • TIF1 beta antibody
    • TIF1-beta antibody
    • TIF1B antibody
    • TIF1B_HUMAN antibody
    • Transcription intermediary factor 1 beta antibody
    • Transcription intermediary factor 1-beta antibody
    • Trim28 antibody
    • Tripartite motif containing 28 antibody
    • tripartite motif containing protein 28 antibody
    • Tripartite motif-containing protein 28 antibody
    see all

画像

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAP1 antibody (ab10483)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAP1 antibody (ab10483)

    Immunohistochemistry analysis of formalin-fixed paraffin embedded sections of human breast tissue staining KAP-1 with ab10483 at 1/1000 dilution, showing the current lot on the left, and previous lot on the right.

  • Immunoprecipitation - Anti-KAP1 antibody (ab10483)
    Immunoprecipitation - Anti-KAP1 antibody (ab10483)

    Immunoprecipitation of KAP-1 from HeLa whole cell lysate using ab10483

    Lane 1: ab10483 current lot

    Lane 2: ab10483 previous lot

    Lane 3: other rabbit anti-KAP-1 antibody

    Lane 4: Control IgG

  • Western blot - Anti-KAP1 antibody (ab10483)
    Western blot - Anti-KAP1 antibody (ab10483)
    All lanes : Anti-KAP1 antibody (ab10483) at 1 µg/ml

    Lane 1 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
    Lane 2 : TCMK-1 (mouse kidney epithelial cell line) whole cell lysate
    Lane 3 : 4T1 (Mouse mammary gland carcinoma cell line) whole cell lysate
    Lane 4 : CT26.WT (murine colon carcinoma) whole cell lysate
    Lane 5 : C6 (rat glioma cell line) whole cell lysate

    Lysates/proteins at 50 µg/ml per lane.

    Predicted band size: 100 kDa


    Exposure time: 3 minutes
  • Western blot - Anti-KAP1 antibody (ab10483)
    Western blot - Anti-KAP1 antibody (ab10483)
    All lanes : Anti-KAP1 antibody (ab10483) at 0.1 µg/ml

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 50 µg
    Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 15 µg
    Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 5 µg
    Lane 4 : HEK293T cells (human epithelial cell line from embryonic kidney transformed with large T antigen) at 50 µg

    Predicted band size: 100 kDa


    Exposure time: 30 seconds
  • Immunoprecipitation - Anti-KAP1 antibody (ab10483)
    Immunoprecipitation - Anti-KAP1 antibody (ab10483)This image was taken from an abreview submitted by an Seth Frietze.
    The ab10483 antibody was used to immunoprecipitate KAP1 from HEK293 nuclear extracts. Three different amounts of ab10483 or control IgG were tested (1, 2 or 4 ug). The eluates from these experiments as well as the supernatants from the KAP1 IPs were analyzed by Western blotting using the ab10483 antibody. As shown by Western blotting, the presence of a ~110 kDa band demonstrated that KAP1 was specifically precipitated from these extracts. The 50 and 25 kDa bands correspond to the IgG of the IP antibodies.

    See Abreview

プロトコール

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

データシートおよび資料

  • SDS download

  • Datasheet download

    Download

参考文献 (59)

ab10483 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab10483 は 59 報の論文で使用されています。

  • Ait-Ammar A  et al. Inhibition of HIV-1 gene transcription by KAP1 in myeloid lineage. Sci Rep 11:2692 (2021). PubMed: 33514850
  • Diao Z  et al. SIRT3 consolidates heterochromatin and counteracts senescence. Nucleic Acids Res 49:4203-4219 (2021). PubMed: 33706382
  • Yu B  et al. B cell-specific XIST complex enforces X-inactivation and restrains atypical B cells. Cell 184:1790-1803.e17 (2021). PubMed: 33735607
  • Wu X  et al. Regulation of cellular sterol homeostasis by the oxygen responsive noncoding RNA lincNORS. Nat Commun 11:4755 (2020). PubMed: 32958772
  • Siebels S  et al. Merkel Cell Polyomavirus DNA Replication Induces Senescence in Human Dermal Fibroblasts in a Kap1/Trim28-Dependent Manner. mBio 11:N/A (2020). PubMed: 32156811
View all Publications for this product

レビューと Q&A

Show All レビュー Q&A
レビューを送る 質問を送る

1-9 of 9 Abreviews or Q&A

ChIP abreview for Anti-KAP1 antibody - ChIP Grade

Poor
Abreviews
Abreviews
abreview image
Application
ChIP
Sample
Human Cell lysate - nuclear (Breast cancer cell line T47D)
Negative control
IgG
Specification
Breast cancer cell line T47D
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: Formaldehid
Positive control
anti H1.2 ab4086
Read More

DR. Albert Jordan

Verified customer

投稿 Apr 09 2019

Western blot abreview for Anti-KAP1 antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293 cells)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-tris)
Loading amount
40 µg
Specification
HEK293 cells
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

投稿 Apr 01 2019

CHIPseq abreview for Anti-KAP1 antibody - ChIP Grade

Poor
Abreviews
Abreviews
abreview image
Application
ChIP-sequencing
Sample
Mouse Cell lysate - nuclear (murine neuronal cell line (Neuro2a))
Specification
murine neuronal cell line (Neuro2a)
Read More

Abcam user community

Verified customer

投稿 Apr 15 2016

Western blot abreview for Anti-KAP1 antibody - ChIP Grade

Below Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (hES and cancer cells)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
100000 cells
Specification
hES and cancer cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
Read More

Abcam user community

Verified customer

投稿 Aug 20 2015

Western blot abreview for Anti-KAP1 antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse embryonic stem cell)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
100000 cells
Specification
mouse embryonic stem cell
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
Read More

Abcam user community

Verified customer

投稿 Aug 20 2015

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-KAP1 antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver, P5)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Na-citrate pH6
Permeabilization
Yes - Triton 0,05%
Specification
Liver, P5
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

投稿 Sep 29 2014

Immunoprecipitation abreview for Anti-KAP1 antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Human Cell lysate - nuclear (Human embryonic kidney cells)
Total protein in input
100 µg
Specification
Human embryonic kidney cells
Immuno-precipitation step
Protein G
Read More

Dr. Seth Frietze

Verified customer

投稿 Jul 04 2011

Question

I have a question about antibodies for ChIP assay from your firm. We used them first in western blot to check their spcificity, and the results are disturbing. We used:
1) H4ac - ab117149 (1:1000)
2) H3K9 - ab117142 (1:1000)
3 ) Kap1 - ab10483 (1:1000)
Pictures in attachment. What can we do to optimize this procedure, and how to be sure if the ChIP reaction with these ab's will be specific.

with regards

Read More

Abcam community

Verified customer

Asked on Oct 01 2012

Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

Read More

Abcam Scientific Support

Answered on Oct 01 2012

Question

I see from your website that synthetic peptides were used to synthesize KAP1 antibodies ab10483 and ab10484. I purchased antibody ab10484 (having apparently overlooked the ChIP-grade ab10483) and have recently tried to use it in a Re-ChIP assay. The background was high for the first IP and the Re-ChIP result was somewhat difficult to interpret because in one context, the signal was much lower than expected and in another it was much higher. Would it be possible to send me information regarding the specific epitopes used in creating these products.

Read More

Abcam community

Verified customer

Asked on Mar 16 2012

Answer

Thank you for contacting us.

While the exact immunogens used for these product are proprietary I am able to share these details about ab10483 and ab10484.

The epitope recognized by AB10483 maps to a region between residue 1 and 50 of human Tripartite Motif-Containing 28 using the numbering given in entry NP_005753.1 (GeneID 10155).
The epitope recognized by AB10484 maps to a region between residue 650 and 700 of human Tripartite Motif-Containing 28 using the numbering given in entry NP_005753.1 (GeneID 10155).

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Abcam Scientific Support

Answered on Mar 16 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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