Validation Data Gallery

Filter: AllWBIPIF/ICC

WB analysis of HeLa using 10456-1-AP
HeLa cells were subjected to SDS PAGE followed by western blot with 10456-1-AP (NELF-A Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
WB analysis using 10456-1-AP
Various lysates were subjected to SDS PAGE followed by western blot with 10456-1-AP (NELF-A antibody) at dilution of 1:8000 incubated at room temperature for 1.5 hours.
WB analysis of mouse brain using 10456-1-AP
mouse brain tissue were subjected to SDS PAGE followed by western blot with 10456-1-AP (NELF-A Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
WB analysis of HeLa using 10456-1-AP
HeLa cells were subjected to SDS PAGE followed by western blot with 10456-1-AP (NELF-A antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.
WB analysis of mouse liver using 10456-1-AP
mouse liver tissue were subjected to SDS PAGE followed by western blot with 10456-1-AP (NELF-A antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.
WB analysis of Jurkat using 10456-1-AP
Jurkat cells were subjected to SDS PAGE followed by western blot with 10456-1-AP (NELF-A antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.
WB analysis of HeLa using 10456-1-AP
HeLa cells were subjected to SDS PAGE followed by western blot with 10456-1-AP (NELF-A Antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.
IP experiment of mouse brain using 10456-1-AP
IP result of anti-NELF-A (IP:10456-1-AP, 4ug; Detection:10456-1-AP 1:500) with mouse brain tissue lysate 3440ug.
IF Staining of HeLa using 10456-1-AP
Immunofluorescent analysis of (4% PFA) fixed HeLa cells using NELF-A antibody (10456-1-AP) at dilution of 1:600 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-phalloidin (red).

Featured Product
KD/KO Validated

NELF-A Polyclonal antibody

NELF-A Polyclonal Antibody for WB, IF/ICC, IP, ELISA

Cat No. 10456-1-AP

Publications(5)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

WB, IF/ICC, IP, ELISA

Synonyms

WHSC2, Wolf-Hirschhorn syndrome candidate 2 protein, NELFA, NELF A

Formulation: PBS and Azide

PBS and Azide

Conjugate: Unconjugated

Unconjugated

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information
Publications (5)
Protocols

データシート［PDF］を印刷

MSDS

Tested Applications

Positive WB detected in	HeLa cells, mouse liver tissue, Jurkat cells, mouse brain tissue
Positive IP detected in	mouse brain tissue
Positive IF/ICC detected in	HeLa 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
Immunofluorescence (IF)/ICC	IF/ICC : 1:300-1:1200
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 2 publications below
WB	See 4 publications below

Product Information

10456-1-AP targets NELF-A in WB, IF/ICC, IP, 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	NELF-A fusion protein Ag0708 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 2
Calculated molecular weight	57 kDa
Observed molecular weight	57 kDa
GenBank accession number	BC002764
Gene Symbol	NELFA
Gene ID (NCBI)	7469
RRID	AB_2216327
Conjugate	Unconjugated
Form	Liquid
Purification Method	Antigen affinity purification
UNIPROT ID	Q9H3P2
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.

Background Information

WHSC2, also known as NELF-A, is a component of the Negative ELongation Factor complex, which inhibits RNA polymerase II progression early during elongation (a process referred to as 'promoter proximal pausing'), thereby altering expression of its target genes in a positive and negative manner [PMID:19245807]. This complex acts with DRB sensitivity-inducing factor (DSIF), a heterodimer of SPT4 and SPT5, to cause transcriptional pausing of RNA polymerase II[PMID:12612062].

Protocols

Product Specific Protocols
WB protocol for NELF-A antibody 10456-1-AP	Download protocol
IF protocol for NELF-A antibody 10456-1-AP	Download protocol
IP protocol for NELF-A antibody 10456-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Publications

Species	Application	Title
		Mol Cell SPT5 stabilization of promoter-proximal RNA polymerase II. Authors - Yuki Aoi View Article
mouse	WB	Nat Commun RNA polymerase II pausing factor NELF in CD8+ T cells promotes antitumor immunity. Authors - Bogang Wu View Article
human	WB	Nucleic Acids Res Multiple P-TEFbs cooperatively regulate the release of promoter-proximally paused RNA polymerase II. Authors - Xiaodong Lu KD Validated View Article
human	WB	Nat Commun ERK-mediated NELF-A phosphorylation promotes transcription elongation of immediate-early genes by releasing promoter-proximal pausing of RNA polymerase II Authors - Seina Ohe KD Validated View Article
human	WB	J Biol Chem An Essential Signaling Function of Cytoplasmic NELFB is Independent of RNA Polymerase II Pausing Authors - Haihui Pan View Article

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

WB analysis of HeLa using 10456-1-AP

WB analysis using 10456-1-AP

WB analysis of mouse brain using 10456-1-AP

WB analysis of HeLa using 10456-1-AP

WB analysis of mouse liver using 10456-1-AP

WB analysis of Jurkat using 10456-1-AP

WB analysis of HeLa using 10456-1-AP

IP experiment of mouse brain using 10456-1-AP

IF Staining of HeLa using 10456-1-AP

NELF-A Polyclonal antibody

NELF-A Polyclonal Antibody for WB, IF/ICC, IP, ELISA

-/ -

Tested Applications

Recommended dilution

Published Applications

Product Information

Background Information

Protocols

Publications

SPT5 stabilization of promoter-proximal RNA polymerase II.

RNA polymerase II pausing factor NELF in CD8+ T cells promotes antitumor immunity.

Multiple P-TEFbs cooperatively regulate the release of promoter-proximally paused RNA polymerase II.

ERK-mediated NELF-A phosphorylation promotes transcription elongation of immediate-early genes by releasing promoter-proximal pausing of RNA polymerase II

An Essential Signaling Function of Cytoplasmic NELFB is Independent of RNA Polymerase II Pausing