SETDB1 Polyclonal antibody

• Featured Product
• KD/KO Validated

SETDB1 Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human and More (1)

Applications

WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ChIP, ELISA

Conjugate

Unconjugated

Publications

78

Reviews (3)

Cat no : 11231-1-AP

Synonyms

Histone-lysine N-methyltransferase SETDB1, Histone H3-K9 methyltransferase 4, H3-K9-HMTase 4, ESET, ERG-associated protein with SET domain

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Validation Data Gallery

• 

  WB analysis using 11231-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 11231-1-AP (SETDB1 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of Jurkat using 11231-1-AP

  WB result of SETDB1 antibody (11231-1-AP, 1:1000) with si-Control and si-SETDB1 transfected Jurkat cells.

• 

  WB analysis of HeLa using 11231-1-AP

  WB result of SETDB1 antibody (11231-1-AP; 1:1000; incubated at room temperature for 1.5 hours) with sh-Control and sh-SETDB1 transfected HeLa cells.

• 

  WB analysis using 11231-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 11231-1-AP (SETDB1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of Jurkat using 11231-1-AP

  Jurkat cells were subjected to SDS PAGE followed by western blot with 11231-1-AP (SETDB1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of human testis using 11231-1-AP

  human testis tissue were subjected to SDS PAGE followed by western blot with 11231-1-AP (SETDB1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of PC-3 using 11231-1-AP

  PC-3 cells were subjected to SDS PAGE followed by western blot with 11231-1-AP (SETDB1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of PC-3 using 11231-1-AP

  PC-3 cells were subjected to SDS PAGE followed by western blot with 11231-1-AP (SETDB1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of MCF-7 using 11231-1-AP

  MCF7 cells were subjected to SDS PAGE followed by western blot with 11231-1-AP (SETDB1 antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.

• 

  WB analysis of MCF-7 using 11231-1-AP

  MCF7 cells were subjected to SDS PAGE followed by western blot with 11231-1-AP (SETDB1 antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HEK-293 using 11231-1-AP

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 11231-1-AP (SETDB1 antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HEK-293 using 11231-1-AP

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 11231-1-AP (SETDB1 antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.

• 

  WB analysis of human testis using 11231-1-AP

  human testis tissue were subjected to SDS PAGE followed by western blot with 11231-1-AP (SETDB1 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of Jurkat using 11231-1-AP

  Jurkat cells were subjected to SDS PAGE followed by western blot with 11231-1-AP (SETDB1 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  IP experiment of MCF-7 using 11231-1-AP

  IP result of anti-SETDB1 (IP:11231-1-AP, 3ug; Detection:11231-1-AP 1:1000) with MCF-7 cells lysate 2500ug.

• 

  IHC staining of human lung cancer using 11231-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 11231-1-AP (SETDB1 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lung cancer using 11231-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 11231-1-AP (SETDB1 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human testis using 11231-1-AP

  Immunohistochemical analysis of paraffin-embedded human testis using 11231-1-AP (SETDB1 antibody) at dilution of 1:50 (under 10x lens).

• 

  IHC staining of human testis using 11231-1-AP

  Immunohistochemical analysis of paraffin-embedded human testis using 11231-1-AP (SETDB1 antibody) at dilution of 1:50 (under 40x lens).

• 

  IHC staining of human testis using 11231-1-AP

  Immunohistochemical analysis of paraffin-embedded human testis using 11231-1-AP (SETDB1 antibody) at dilution of 1:50 (under 10x lens).

• 

  IHC staining of human testis using 11231-1-AP

  Immunohistochemical analysis of paraffin-embedded human testis using 11231-1-AP (SETDB1 antibody) at dilution of 1:50 (under 40x lens).

• 

  IF Staining of A431 using 11231-1-AP

  Immunofluorescent analysis of (4% PFA) fixed A431 cells using SETDB1 antibody (11231-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L).

• 

  FC experiment of A431 using 11231-1-AP

  1x10^6 A431 cells were intracellularly stained with 0.4 ug SETDB1 Polyclonal antibody (11231-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.4 ug Rabbit IgG control Rabbit PolyAb (30000-0-AP) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: Jurkat cells, HEK-293 cells, HeLa cells, human testis tissue, MCF-7 cells, PC-3 cells
• Positive IP detected in: MCF-7 cells
• Positive IHC detected in: human lung cancer tissue, human testis 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
• Positive FC (Intra) detected in: A431 cells

Recommended dilution

• Western Blot (WB): WB : 1:500-1:2000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

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.

Published Applications

• KD/KO: See 21 publications below
• WB: See 59 publications below
• IHC: See 7 publications below
• IF: See 11 publications below
• IP: See 9 publications below
• CoIP: See 3 publications below
• ChIP: See 22 publications below

Product Information

11231-1-AP targets SETDB1 in WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ChIP, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Cited Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: SETDB1 fusion protein Ag1725
• Full Name: SET domain, bifurcated 1
• Calculated Molecular Weight: 143 kDa
• Observed Molecular Weight: 170 kDa
• GenBank Accession Number: BC009362
• Gene Symbol: SETDB1
• Gene ID (NCBI): 9869
• RRID: AB_2186069
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity 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. This antibody is a rabbit polyclonal antibody raised against residues near the N terminus of human SETDB1.The calculated molecular weight of SETDB1 is 143 kDa, but the modified SETDB1 protein is about 170 kDa (PMID: 11791185).

Publications

Species	Application	Title
mouse	WB	Nature; METTL3 regulates heterochromatin in mouse embryonic stem cells.; Authors - Wenqi Xu ; KO Validated
mouse	WB,CoIP	Nature; The RNA m6A reader YTHDC1 silences retrotransposons and guards ES cell identity.; Authors - Jiadong Liu
mouse,human	WB	Nature; Epigenetic silencing by SETDB1 suppresses tumour intrinsic immunogenicity.; Authors - Gabriel K Griffin ; KO Validated
mouse	WB,CoIP,ChIP	Nature; KDM5B promotes immune evasion by recruiting SETDB1 to silence retroelements; Authors - Shang-Min Zhang
human	WB	Nature; Genome surveillance by HUSH-mediated silencing of intronless mobile elements.; Authors - Marta Seczynska ; KD Validated
human	WB	Science; GENE SILENCING. Epigenetic silencing by the HUSH complex mediates position-effect variegation in human cells.; Authors - Iva A Tchasovnikarova ; KD Validated

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Hilda (Verified Customer) (07-29-2024)

SETDB1 antibody works very well with human and mouse cell lines for both western blot and ChIP. It requires a short incubation period and produces distinct bands, with a few nonspecific bands observed variable to the type of cells used. CHIP outcomes are still preliminary, but it has resulted in good binding at 2ug antibody used.

• Applications: Western Blot, ChIP
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: 4T1 Cells

SETDB1 Antibody Western Blot,ChIP validation (1:1000 dilution) in 4T1 Cells (Cat no:11231-1-AP)

FH

Roy (Verified Customer) (06-28-2024)

Does work well to detect SETDB1 in Jurkat and HeLa cells (Overnight incubation at 4°C, 1/1000 dilution)

• Applications: Western Blot
• Primary Antibody Dilution: 1/1000
• Cell Tissue Type: Jurkat/HeLa

FH

Taku (Verified Customer) (06-15-2022)

• Applications: Western Blot
• Primary Antibody Dilution: x2000
• Cell Tissue Type: T cell

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