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KD/KO Validated

SETDB1 Polyclonal antibody

Name: SETDB1 Polyclonal antibody
Brand: Proteintech
SKU: 11231-1-AP
Rating: 4.67 (3 reviews)

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

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

Validation Data Gallery

Filter:

AllWBIPIHCIF/ICCFC (Intra)

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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Cat No. 11231-1-AP

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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

Application	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).

Protocols

Product Specific Protocols
WB protocol for SETDB1 antibody 11231-1-AP	Download protocol
IHC protocol for SETDB1 antibody 11231-1-AP	Download protocol
IF protocol for SETDB1 antibody 11231-1-AP	Download protocol
IP protocol for SETDB1 antibody 11231-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Publications

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

Reviews

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

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

SETDB1 Polyclonal antibody

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

Validation Data Gallery

WB analysis using 11231-1-AP

WB analysis of Jurkat using 11231-1-AP

WB analysis of HeLa using 11231-1-AP

WB analysis using 11231-1-AP

WB analysis of Jurkat using 11231-1-AP

WB analysis of human testis using 11231-1-AP

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

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

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

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

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

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

WB analysis of human testis using 11231-1-AP

WB analysis of Jurkat using 11231-1-AP

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

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

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

IHC staining of human testis using 11231-1-AP

IHC staining of human testis using 11231-1-AP

IHC staining of human testis using 11231-1-AP

IHC staining of human testis using 11231-1-AP

IF Staining of A431 using 11231-1-AP

FC experiment of A431 using 11231-1-AP

Tested Applications

Recommended dilution

Published Applications

Product Information

Background Information

Protocols

Publications

METTL3 regulates heterochromatin in mouse embryonic stem cells.

The RNA m6A reader YTHDC1 silences retrotransposons and guards ES cell identity.

Epigenetic silencing by SETDB1 suppresses tumour intrinsic immunogenicity.

KDM5B promotes immune evasion by recruiting SETDB1 to silence retroelements

Genome surveillance by HUSH-mediated silencing of intronless mobile elements.

GENE SILENCING. Epigenetic silencing by the HUSH complex mediates position-effect variegation in human cells.

Reviews

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