Validation Data Gallery

Filter: AllWBIPIHCIF/ICCFC (Intra)ChIP

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 1:6000 incubated at room temperature for 1.5 hours.
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 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 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.
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 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.
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.
IP experiment of MCF-7 using 11231-1-AP
IP result of anti-SETDB1 (IP:11231-1-AP, 4ug; Detection:11231-1-AP 1:3000) with MCF-7 cells lysate 1720 ug.
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 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).
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).
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 AffiniPure 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 AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.4 ug rabbit IgG isotype control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer.
ChIP experiment of HeLa using 11231-1-AP
Chromatin was prepared from HeLa cells, cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 15 µg of cross-linked chromatin, 5 µg of SETDB1 Antibody (11231-1-AP) or 5 ug of Normal Rabbit IgG (30000-0-AP), and 30 µl of Protein A Magarose Beads. The immunoprecipitated DNA was quantified by real time PCR. Primers are located in the first kb of the transcribed region.

Featured Product
KD/KO Validated

SETDB1 Polyclonal antibody

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

Cat No. 11231-1-AP

Publications(93)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

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

Synonyms

EC:2.1.1.366, ERG-associated protein with SET domain, ESET, H3-K9-HMTase 4, Histone H3-K9 methyltransferase 4

Formulation: PBS, Azide, Glycerol

PBS, Azide, Glycerol

Conjugate: Unconjugated

Unconjugated

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information
Publications (93)
Protocols

データシート［PDF］を印刷

MSDS

Tested Applications

Positive WB detected in	HEK-293 cells, HeLa cells, human testis tissue, Jurkat cells, MCF-7 cells, PC-3 cells, MDA-MB-231 cells, NIH/3T3 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
Positive ChIP detected in	HeLa cells

Recommended dilution

Application	Dilution
Western Blot (WB)	WB : 1:2000-1:12000
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
Chromatin immunoprecipitation (ChIP)	CHIP : 1:10-1:100
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 68 publications below
IHC	See 10 publications below
IF	See 12 publications below
IP	See 9 publications below
CoIP	See 4 publications below
ChIP	See 24 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, mouse samples.

Tested Reactivity	human, mouse
Cited Reactivity	human, mouse
Host / Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	CatNo: Ag1725 Product name: Recombinant human SETDB1 protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: 1-350 aa of BC009362 Sequence: MSSLPGCIGLDAATATVESEEIAELQQAVVEELGISMEELRHFIDEELEKMDCVQQRKKQLAELETWVIQKESEVAHVDQLFDDASRAVTNCESLVKDFYSKLGLQYRDSSSEDESSRPTEIIEIPDEDDDVLSIDSGDAGSRTPKDQKLREAMAALRKSAQDVQKFMDAVNKKSSSQDLHKGTLSQMSGELSKDGDLIVSMRILGKKRTKTWHKGTLIAIQTVGPGKKYKVKFDNKGKSLLSGNHIAYDYHPPADKLYVGSRVVAKYKDGNQVWLYAGIVAETPNVKNKLRFLIFFDDGYASYVTQSELYPICRPLKKTWEDIEDISCRDFIEEYVTAYPNRPMVLLKS 相同性解析による交差性が予測される生物種
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
Form	Liquid
Purification Method	Antigen affinity purification
UNIPROT ID	Q15047
Storage Buffer	PBS with 0.02% sodium azide and 50% glycerol{{ptg:BufferTemp}}7.3
Storage Conditions	Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

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

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

WB analysis of Jurkat using 11231-1-AP

WB analysis of HeLa using 11231-1-AP

WB analysis using 11231-1-AP

WB analysis 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 human testis using 11231-1-AP

WB analysis of Jurkat 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 Jurkat using 11231-1-AP

WB analysis of human testis using 11231-1-AP

IP experiment of MCF-7 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 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

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

IF Staining of A431 using 11231-1-AP

FC experiment of A431 using 11231-1-AP

ChIP experiment of HeLa using 11231-1-AP

SETDB1 Polyclonal antibody

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

-/ -

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.