Anti-p-KAP-1 (Ser824) Antibody, clone 1K9 ZooMAb^® Rabbit Monoclonal

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ZooMAb^® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb^® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb^® antibodies are reliably available and ready to ship when you need them.

Clone 1K9 is a ZooMAb^® rabbit recombinant monoclonal antibody that specifically detects KAP-1/TRIM28 phosphorylated on serine 824.

KLH-conjugated linear peptide corresponding to 11 amino acids surrounding phosphoserine 824 from the C-terminal region of human KAP-1/TRIM28.

Quality Control Testing

Evaluated by Western Blotting in lysate from HeLa cells treated with Etoposide.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Phospho-KAP-1 (Ser824) in lysate from HeLa cells treated with Etoposide (20 g/mL; 3 h).

Quality Control Testing

Evaluated by Western Blotting in lysate from HeLa cells treated with Etoposide.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Phospho-KAP-1 (Ser824) in lysate from HeLa cells treated with Etoposide (20 g/mL; 3 h).

Tested Applications

Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected Phospho-KAP-1 (Ser824) in HeLa cells treated with and without etoposide.

Peptide Inhibition Assay: Target band detection in lysate from HeLa cells treated with Etoposide was prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide.

Affinity Binding Assay: A representative lot of this antibody bound p-KAP-1(Ser824) peptide with a KD of 8.5 x 10-8 in an affinity binding assay.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Transcription intermediary factor 1-beta (UniProt Q13263; also known as EC:2.3.2.27, E3 SUMO-protein ligase TRIM28, KAP-1, KRAB-associated protein 1, KRAB-interacting protein 1, KRIP-1, RING finger protein 96, TIF1-beta, Transcriptional intermediary factor 1-beta, Tripartite motif-containing 28) is encoded by the TRIM28 (also known as KAP1, PPP1R157, RNF96, TF1B, TIF1B) gene (Gene ID: 10155) in human. KAP-1 is a widely expressed, oligomeric, NuCLEAR^™ corepressor for KRAB domain-containing zinc finger protein that mediates gene silencing by recruiting chromodomain helicase DNA binding protein 3 (CHD3) complex and SET domain bifurcated histone lysine methyltransferase 1 (SETDB1) to the promoter regions of KRAB target genes. It enhances transcriptional repression by coordinating the increase in H3K9me, the decrease in histone H3 lysine 9 and lysine 14 acetylation, and the disposition of HP1 proteins to Silence | Speed | Simplicity^®| Speed | Simplicity"]gene expression. Its PHD-type zinc finger (aa 625-672) enhances CCAAT enhancer binding protein β (CEBPB) transcriptional activity. The PHD-type zinc finger, the HP1 box (aa 476-513) and the bromo domain (aa 697-801) are reported to function together to assemble the machinery required for repression of KRAB domain-containing proteins. The RING-finger-B Box-coiled-coil/tripartite motif (RBCC/TRIM motif; aa 65-376) is required for interaction with the KRAB domain of KRAB-zinc finger proteins. KAP-1 undergoes phosphorylation by ATM on serine 824 that 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 PPP1CA and PP1CB allows sumoylation and expression of TRIM28 target genes. This ZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, Affinity^™, reproducibility, and stability over conventional monoclonals. (Ref.: Li, X., et al. (2007). J. Biol. Chem. 282(50); 36177-36189; Ivanov, AV., et al. (2007). Mol. Cell. 28(5); 823-837; Ziv, Y., et al. (2006). Nat. Cell Biol. 8(8); 870-876).

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.

300 µg/mL after reconstitution at 25 µL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Affinity is a trademark of Mine Safety Appliances Co.

NuCLEAR is a trademark of Sigma-Aldrich Co. LLC

Silence | Speed | Simplicity is a registered trademark of Eppendorf AG

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.