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Immunoprecipitation - KAP1 antibody (ab10484)
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-KAP1 antibody(ab10484)
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Immunocytochemistry/ Immunofluorescence-KAP1 antibody(ab10484)
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Product Name
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KAP1 antibody
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See all KAP1 antibodies (14)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to KAP1
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Immunogen
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Immunogen was a synthetic peptide, which represented a portion of human tripartite motif-containing 28 (KAP1) encoded within exon 14 (LocusLink ID 10155).
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Reacts with
(species key)
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Hu, Ms
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Tested applications
(see key)
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ICC/IF, IHC-P, IP, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
IHC-P: Use at a concentration of 2 µg/ml. See also PMID 19086899.
ICC/IF: Use at a concentration of 1 µg/ml.
IP: Use at a concentration of 1 - 5 µg/ml.
WB: 1/500 - 1/2000. Detects a band of approximately 110 kDa (predicted molecular weight: 100 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
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WB: Whole cell lysate from normal 293T cells or 293T cells transiently transfected with a human KAP1 construct, embryonic stem cell lysate (see Abreview)
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Cellular localization
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Nuclear
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Research areas
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Chromatin and Nuclear Signaling >> Transcription >> Transcription Factors
Chromatin and Nuclear Signaling >> Transcription >> Other factors
Cell Biology >> Proteolysis / Ubiquitin >> Proteasome / Ubiquitin >> Ubiquitin E2s and E3s >> RING Finger E3 Ligase
Chromatin and Nuclear Signaling >> Chromatin Binding Proteins >> Methylated DNA
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Relevance
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The KRAB (Kruppel Associated Box) domain is minimally about 45 amino acids in length and is a transcriptional repression domain found in numerous transcription factors. Over 220 KRAB zinc finger protein (KRAB ZFP) genes have been identified in the human genome. These proteins functionally repress transcription via specific interactions with KAP1 (KRAB Associated Protein 1).
KAP1 is an 835 amino acid polypeptide that contains a RING finger, B boxes, and a PHD finger. KAP1 has been shown to form complexes with KRAB domain transcription factors and increase the efficiency with which they mediate repression. KAP1 has also been shown to directly interact with HP1 (heterochromatin protein 1) and KRAZ1 (Kruppel associated box containing zinc finger protein 1). KAP1 directly targets KRAZ1 to the foci of centromeric heterochromatin containing HP1alpha, helping to regulate transcriptional repression. Recent studies have shown that KAP1 mutants with the ability to bind KRAB but unable to bind HP1 leads to random distribution of KRAZ1 and strong transcriptional activation.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.1% Sodium Azide
Constituents: PBS, 8mM Phosphate, 60mM Citrate, 150mM Tris, pH 7-8 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Purification notes
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Antibodies were affinity purified using the peptide immobilized on solid support.
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Form
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Liquid
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Concentration
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1.000 mg/ml
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Storage instructions
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Store at +4°C.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this KAP1 antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for KAP1 antibody images, references, products related to ab10484 and other tools.
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KAP1 antibody images:
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 Immunoprecipitation - KAP1 antibody (ab10484)
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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-KAP1 antibody(ab10484)
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.
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Immunocytochemistry/ Immunofluorescence-KAP1 antibody(ab10484)
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.
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References for KAP1 antibody (ab10484)
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This product has been used in: (two most recent references)
Hu G et al. A genome-wide RNAi screen identifies a new transcriptional module required for self-renewal. Genes Dev 23:837-48 (2009). WB; Mouse. PubMed: 19339689
Ho J et al. Novel breast cancer metastasis-associated proteins. J Proteome Res 8:583-94 (2009). WB, IHC-P; Human. PubMed: 19086899
If you publish research using ab10484 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to KAP1
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KAP1 antibody - more information
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