SETDB1 Monoclonal antibody

SETDB1 Monoclonal Antibody for WB, IHC, IF/ICC, ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, ELISA

Conjugate

Unconjugated

CloneNo.

1H6E5

Cat no : 66293-1-Ig

Synonyms

ESET, H3 K9 HMTase 4, H3 K9 HMTase4, KG1T, KIAA0067, KMT1E, Lysine N methyltransferase 1E, SET domain bifurcated 1, SET domain, bifurcated 1, SETDB1

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Validation Data Gallery

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  WB analysis using 66293-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66293-1-Ig (SETDB1 antibody) at dilution of 1:100000 incubated at room temperature for 1.5 hours.

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  WB analysis of human heart using 66293-1-Ig

  human heart tissue were subjected to SDS PAGE followed by western blot with 66293-1-Ig (SETDB1 Antibody) at dilution of 1:8000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 66293-1-Ig

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 66293-1-Ig (SETDB1 Antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  IHC staining of mouse colon using 66293-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse colon tissue slide using 66293-1-Ig (SETDB1 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human colon using 66293-1-Ig

  Immunohistochemical analysis of paraffin-embedded human colon tissue slide using 66293-1-Ig (SETDB1 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat liver using 66293-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat liver tissue slide using 66293-1-Ig (SETDB1 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat colon using 66293-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat colon tissue slide using 66293-1-Ig (SETDB1 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human colon using 66293-1-Ig

  Immunohistochemical analysis of paraffin-embedded human colon tissue slide using 66293-1-Ig (SETDB1 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human colon using 66293-1-Ig

  Immunohistochemical analysis of paraffin-embedded human colon tissue slide using 66293-1-Ig (SETDB1 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of A431 using 66293-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed A431 cells using SETDB1 antibody (66293-1-Ig, Clone: 1H6E5 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

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Tested Applications

• Positive WB detected in: HeLa cells, HEK-293 cells, human heart tissue, MCF-7 cells, Jurkat cells, HepG2 cells
• Positive IHC detected in: human colon tissue, rat liver tissue, rat colon tissue, mouse colon tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: A431 cells

Recommended dilution

• Western Blot (WB): WB : 1:20000-1:100000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

66293-1-Ig targets SETDB1 in WB, IHC, IF/ICC, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: SETDB1 fusion protein Ag21644
• Full Name: SET domain, bifurcated 1
• Calculated Molecular Weight: 143 kDa
• Observed Molecular Weight: 170-180 kDa
• GenBank Accession Number: BC009362
• Gene Symbol: SETDB1
• Gene ID (NCBI): 9869
• RRID: AB_2881676
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

SETDB1, also named as ESET, KIAA0067 and KMT1E, belongs to the histone-lysine methyltransferase family. It is a SET domain protein with histone H3-K9-specific methyltransferase activity. H3 'Lys-9' trimethylation is coordinated with DNA methylation and represents a specific tag for epigenetic transcriptional repression by recruiting HP1 (CBX1, CBX3 and/or CBX5) proteins to methylated histones. SETDB1 mainly functions in euchromatin regions, thereby playing a central role in the silencing of euchromatic genes. It 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 regulates histone methylation, gene silencing, and transcriptional repression. It has been identified as a target for treatment in Huntington Disease, given that gene silencing and transcription dysfunction likely play a role in the disease pathogenesis. The calculated molecular weight of SETDB1 is 143 kDa, but the modified SETDB1 protein is about 170 kDa (PMID: 11791185).