概述

  • 产品名称Anti-KAP1抗体
    参阅全部 KAP1 一抗
  • 描述
    兔多克隆抗体to KAP1
  • 经测试应用适用于: WB, IP, ICC/IF, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Human
    预测可用于: Rat, Rabbit, Chimpanzee, Rhesus monkey, Gorilla
  • 免疫原

    Immunogen was a synthetic peptide, which represented a portion of human tripartite motif-containing 28 (KAP1) encoded within exon 14 (LocusLink ID 10155).

  • 阳性对照

性能

应用

Our Abpromise guarantee covers the use of ab10484 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 1/500 - 1/10000. Detects a band of approximately 110 kDa (predicted molecular weight: 100 kDa).
IP Use at 2-10 µg/mg of lysate.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 2 µg/ml.

靶标

  • 功能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.
  • 组织特异性Expressed in all tissues tested including spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes.
  • 通路Protein modification; protein sumoylation.
  • 序列相似性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.
  • 结构域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.
  • 翻译后修饰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.
  • 细胞定位Nucleus. Associated with centromeric heterochromatin during cell differentiation through CBX1.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-KAP1 antibody 图像

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma (left) and ovarian carcinoma (right) tissues labelling KAP1 with ab10484 at 1/5000 (0.2µg/ml) and 1/1000 (1µg/ml). Detection: DAB.
  • Detection of Human KAP1 by Western Blot and Immunoprecipitation. Samples: A. Whole cell lysate (50 ug) from normal 293T cells (E) or 293T cells transiently transfected with a human KAP1 construct (T). B. Whole cell lysate (0.5 mg) from normal 293T cells. Antibodies: ab10484 used at 1 ug/ml for WB (A) or 2 ug/0.5 mg lysate for IP (B). Immunoprecipitated KAP1 was detected using a KAP1 antibody from another source. Detection: Chemiluminescence with a 10 minute (A) or 1 minute (B) exposure.

  • Ab10484 staining normal human spleen. Staining is localized to the nucleus.
    Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • ICC/IF image of ab10484 stained HeLa cells. The cells were 4% PFA 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 (ab10484, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.

  • ab10484 staining KAP1 in murine coronal brain tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with paraformaldehyde and permeabilized using 1% Triton. Samples were then blocked with 5% serum for 1 hour at 25°C followed by incubation with the primary antibody at 2µg/ml for 16 hours at 4°C. An Alexa-Fluor 488-conjugated donkey anti-rabbit polyclonal was used as secondary antibody at a 1/1000 dilution.
    Dentate gyrus of the hippocampus immunostained with DAPI (top) and KAP1 (bottom), which nicely labels the nuclei of the entire dentate gyrus.

Anti-KAP1 antibody (ab10484)参考文献

This product has been referenced in:

See all 10 Publications for this product

Product Wall

Thank you for calling us and for alerting us to the problem you are experiencing with our product. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

As we discussed on the ph...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (U2oS)
Specification U2oS
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Tween
Blocking step BSA as blocking agent for 45 minute(s) · Concentration: 2% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Aug 07 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (smooth muscle cells)
Loading amount 25 µg
Specification smooth muscle cells
Gel Running Conditions Reduced Denaturing (8)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Yiwen Liu

Verified customer

提交于 Apr 16 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - other (U2OS)
Loading amount 20 µg
Specification U2OS
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Dr. Andrew Cobb

Verified customer

提交于 Apr 02 2012

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 o...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Coronal Brain Section)
Specification Coronal Brain Section
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization Yes - 1% Triton
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Apr 25 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - whole cell (HeLa Cells)
Total protein in input 3 µg
Specification HeLa Cells
Immuno-precipitation step Protein A
Username

Abcam user community

Verified customer

提交于 Aug 25 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (embryonic stem cells)
Specification embryonic stem cells
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5%
Username

Dr. M Pardo

Verified customer

提交于 Nov 16 2005

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"