Anti-KMT1E / SETDB1 Rabbit mAb

Product

Anti-KMT1E / SETDB1 Rabbit mAb

Catalog number :AT0409

Histone methyltransferase that specifically trimethylates 'Lys-9' of histone H3. H3 'Lys-9' trimethylation represents a specific tag for epigenetic transcriptional repression by recruiting HP1 (CBX1, CBX3 and/or CBX5) proteins to methylated histones. Mainly functions in euchromatin regions, thereby playing a central role in the silencing of euchromatic genes. H3 'Lys-9' trimethylation is coordinated with DNA methylation. Probably forms a complex with MBD1 and ATF7IP that represses transcription and couples DNA methylation and histone 'Lys-9' trimethylation. Its activity is dependent on MBD1 and is heritably maintained through DNA replication by being recruited by CAF-1. SETDB1 is targeted to histone H3 by TRIM28/TIF1B, a factor recruited by KRAB zinc-finger proteins.

Overview

Description: Rabbit monoclonal antibody to KMT1E / SETDB1

Reactivity: Mouse, Human, Rat

Tested applications: WB: 1/1000 - 1/10000. Predicted molecular weight: 155 kDa.

IHC-P: Use at an assay dependent dilution.

ICC/IF: Use at an assay dependent concentration. PubMed: 21921037

IP: Use a concentration of 1 - 4 mg/ml.

Properties

Immunogen: Immunogen was a synthetic peptide, which represented a portion of human SET domain, bifurcated 1 encoded within exon 19 (LocusLink ID 9869).

Clonality: Monoclonal, clone number: 1F7

Isotype: Rabbit IgG

Form: Liquid
Preservative: 0.1% Sodium Azide
Constituents: 8mM PBS, 60mM Citrate, 150mM Tris, pH 7-8

Storage instruction: Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.

Database links

Applications

IHC Image: AT0409 (4ug/ml) staining KMT1E in human testis using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of KMT1E in nuclear compartments of the germinal epithelium.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min 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, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

IP/CoIP Image: Whole cell lysate (2 mg) from 293T cells that were mock transfected (1 and 2) or transfected with a SETDB1 expression construct (3 and 4). Rabbit anti-SETDB1 antibodies; AT0409 (2 and 4) and a competitor's antiserum (1 and 3) were used at 1 mcg/mg lysate for IP. AT0409 was used at 0.2 mg/ml for Western Blot Detection: Chemiluminescence with exposure times of 1 to 5 minutes.

Anti-KMT1E / SETDB1 Rabbit mAb

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