Validation Data Gallery

Filter: AllWBIPIHCIF/ICCFC (Intra)

WB analysis of HeLa using 11345-1-AP
HeLa cells were subjected to SDS PAGE followed by western blot with 11345-1-AP (NSD3 antibody at dilution of 1:600 incubated at room temperature for 1.5 hours.
WB analysis using 11345-1-AP
Various lysates were subjected to SDS PAGE followed by western blot with 11345-1-AP (NSD3 antibody) at dilution of 1:1500 incubated at room temperature for 1.5 hours.
WB analysis of rat heart using 11345-1-AP
rat heart tissue were subjected to SDS PAGE followed by western blot with 11345-1-AP (NSD3 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.
WB analysis of HEK-293 using 11345-1-AP
HEK-293 cells were subjected to SDS PAGE followed by western blot with 11345-1-AP (NSD3 antibody at dilution of 1:600 incubated at room temperature for 1.5 hours.
IP experiment of HeLa using 11345-1-AP
IP result of anti-NSD3 (IP:11345-1-AP, 4ug; Detection:11345-1-AP 1:600) with HeLa cells lysate 1085 ug.
IHC staining of human lung squamous cell carcinoma using 11345-1-AP
Immunohistochemical analysis of paraffin-embedded human lung squamous cell carcinoma tissue slide using 11345-1-AP (NSD3 antibody) at dilution of 1:800 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IF Staining of MCF-7 using 11345-1-AP
Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using NSD3 antibody (11345-1-AP) at dilution of 1:400 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002).
FC experiment of NIH/3T3 using 11345-1-AP
1x10^6 NIH/3T3 cells were intracellularly stained with 0.25 ug NSD3 Polyclonal antibody (11345-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.25 ug rabbit IgG isotype control (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

Featured Product
KD/KO Validated

NSD3 Polyclonal antibody

NSD3 Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Cat No. 11345-1-AP

Publications(17)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

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

Synonyms

WHSC1L1, DC28, EC:2.1.1.370, Histone-lysine N-methyltransferase NSD3, Nuclear SET domain-containing protein 3

Formulation: PBS, Azide, Glycerol

PBS, Azide, Glycerol

Conjugate: Unconjugated

Unconjugated

CoraLite® Plus 488 CoraLite® Plus 647

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information
Publications (17)
Protocols

データシート［PDF］を印刷

MSDS

Tested Applications

Positive WB detected in	HeLa cells, HEK-293 cells, rat heart tissue, mouse heart tissue
Positive IP detected in	HeLa cells
Positive IHC detected in	human lung squamous cell carcinoma 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	MCF-7 cells
Positive FC (Intra) detected in	NIH/3T3 cells

Recommended dilution

Application	Dilution
Western Blot (WB)	WB : 1:500-1:1000
Immunoprecipitation (IP)	IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
Immunohistochemistry (IHC)	IHC : 1:400-1:1600
Immunofluorescence (IF)/ICC	IF/ICC : 1:200-1:800
Flow Cytometry (FC) (INTRA)	FC (INTRA) : 0.25 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 3 publications below
WB	See 11 publications below
IHC	See 6 publications below
IF	See 2 publications below
ChIP	See 1 publications below

Product Information

11345-1-AP targets NSD3 in WB, IHC, IF/ICC, FC (Intra), IP, chIP, ELISA applications and shows reactivity with human, mouse, rat samples.

Tested Reactivity	human, mouse, rat
Cited Reactivity	human
Host / Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	CatNo: Ag1894 Product name: Recombinant human WHSC1L1 protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: 1-300 aa of BC012059 Sequence: MDFSFSFMQGIMGNTIQQPPQLIDSANIRQEDAFDNNSDIAEDGGQTPYEATLQQGFQYPATTEDLPPLTNGYPSSISVYETQTKYQSYNQYPNGSANGFGAVRNFSPTDYYHSEIPNTRPHEILEKPSPPQPPPPPSVPQTVIPKKTGSPEIKLKITKTIQNGRELFESSLCGDLLNEVQASEHTKSKHESRKEKRKKSNKHDSSRSEERKSHKIPKLEPEEQNRPNERVDTVSEKPREEPVLKEEAPVQPILSSVPTTEVSTGVKFQVGDLVWSKVGTYPWWPCMVSSDPQLEVHTKI 相同性解析による交差性が予測される生物種
Full Name	Wolf-Hirschhorn syndrome candidate 1-like 1
Calculated molecular weight	1437 aa, 162 kDa
Observed molecular weight	160-170 kDa, 68-72 kDa
GenBank accession number	BC012059
Gene Symbol	WHSC1L1
Gene ID (NCBI)	54904
RRID	AB_2216189
Conjugate	Unconjugated
Form
Form	Liquid
Purification Method	Antigen affinity purification
UNIPROT ID	Q9BZ95
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

WHSC1L1, also named as NSD3 and DC28, belongs to the histone-lysine methyltransferase family and SET2 subfamily. It is histone methyltransferase. WHSC1L1 is a potently transforming oncogenes based on the number of altered phenotypes expressed by the cells. It contains a PWWP-domain that is a methyl-lysine recognition motif involved in histone code modification and epigenetic regulation of gene expression. Knockdown of WHSC1L1 in 8p11-12-amplified breast cancer cells resulted in profound loss of growth and survival of these cells. (PMID: 20940404). This antibody recognize the long isoform (160-170 kDa) and short isoform (68-72 kDa).

Protocols

Product Specific Protocols
IF protocol for NSD3 antibody 11345-1-AP	Download protocol
IHC protocol for NSD3 antibody 11345-1-AP	Download protocol
IP protocol for NSD3 antibody 11345-1-AP	Download protocol
WB protocol for NSD3 antibody 11345-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Publications

Species	Application	Title
human	WB	Sci Adv Arginine methylation of BRD4 by PRMT2/4 governs transcription and DNA repair Authors - Liu Liu View Article
human	IHC	Cancer Res NSD3-Induced Methylation of H3K36 Activates NOTCH Signaling to Drive Breast Tumor Initiation and Metastatic Progression. Authors - Ga-Young Jeong View Article
human	WB	Cancer Res Transforming properties of 8p11-12 amplified genes in human breast cancer. Authors - Yang Zeng-Quan ZQ KD Validated View Article
		Mol Oncol Amplification of WHSC1L1 regulates expression and estrogen-independent activation of ERα in SUM-44 breast cancer cells and is associated with ERα over-expression in breast cancer. Authors - Jonathan C Irish View Article
		Sci Rep NSD2 contributes to oncogenic RAS-driven transcription in lung cancer cells through long-range epigenetic activation. Authors - Verónica García-Carpizo View Article
human	WB,IF	Sci Rep WHSC1L1-mediated EGFR mono-methylation enhances the cytoplasmic and nuclear oncogenic activity of EGFR in head and neck cancer. Authors - Vassiliki Saloura KD Validated View Article

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

WB analysis of HeLa using 11345-1-AP

WB analysis using 11345-1-AP

WB analysis of rat heart using 11345-1-AP

WB analysis of HEK-293 using 11345-1-AP

IP experiment of HeLa using 11345-1-AP

IHC staining of human lung squamous cell carcinoma using 11345-1-AP

IF Staining of MCF-7 using 11345-1-AP

FC experiment of NIH/3T3 using 11345-1-AP

NSD3 Polyclonal antibody

NSD3 Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

-/ -

Tested Applications

Recommended dilution

Published Applications

Product Information

Background Information

Protocols

Publications

Arginine methylation of BRD4 by PRMT2/4 governs transcription and DNA repair

NSD3-Induced Methylation of H3K36 Activates NOTCH Signaling to Drive Breast Tumor Initiation and Metastatic Progression.

Transforming properties of 8p11-12 amplified genes in human breast cancer.

Amplification of WHSC1L1 regulates expression and estrogen-independent activation of ERα in SUM-44 breast cancer cells and is associated with ERα over-expression in breast cancer.

NSD2 contributes to oncogenic RAS-driven transcription in lung cancer cells through long-range epigenetic activation.

WHSC1L1-mediated EGFR mono-methylation enhances the cytoplasmic and nuclear oncogenic activity of EGFR in head and neck cancer.