Phospho-H2AX (Ser139) Polyclonal Antibody

Formalin-fixed and paraffin embedded human bladder carcinoma labeled with Anti-Phospho-Histone H2A.X (Ser139) Polyclonal Antibody, Unconjugated (bs-3185R) at 1:200 followed by conjugation to the secondary antibody and DAB staining.

Hela cell lysates probed with Histone H2A.X (Ser139) Polyclonal Antibody, Unconjugated (bs-3185R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

$Hela cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Histone H2A.X (Ser140) Antibody(bs-3185R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

Hela cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Histone H2A.X (Ser140) Antibody(bs-3185R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

$Lane 1: Mouse BV2 cell lysates; Lane 2: Rat Testis tissue lysates; Lane 3: Human 293T cell lysates; Lane 4: Human Jurkat cell lysates probed with Phospho-H2AX (Ser139) Polyclonal Antibody, Unconjugated (bs-3185R) at 1:5000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.$

Lane 1: Mouse BV2 cell lysates; Lane 2: Rat Testis tissue lysates; Lane 3: Human 293T cell lysates; Lane 4: Human Jurkat cell lysates probed with Phospho-H2AX (Ser139) Polyclonal Antibody, Unconjugated (bs-3185R) at 1:5000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

Applications

WB
ELISA
FCM
IHC-P
IHC-F
IF(IHC-P)
IF(IHC-F)
IF(ICC)

Reactivity

Human
Mouse
Rat

Predicted Reactivity

Dog
Cow
Pig
Horse
Rabbit

Overview
Catalog #	bs-3185R
Product Name	Phospho-H2AX (Ser139) Polyclonal Antibody
Applications	WB ELISA FCM IHC-P IHC-F IF(IHC-P) IF(IHC-F) IF(ICC)
Specificity	This phosphorylation site is homologous to that of Ser140 in Mouse and Rat.
Reactivity	Human, Mouse, Rat
Predicted Reactivity	Dog, Cow, Pig, Horse, Rabbit
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic phosphopeptide derived from human Histone H2AX around the phosphorylation site of Ser140
Modification Site	Ser140
Clonality	Polyclonal
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4_. Store at -20_ for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	3014
Swiss Prot	P16104
Subcellular location	Nucleus
Synonyms	H2AX; H2A.X; H2A/X; Histone H2AX; Histone H2A.X; H2AFX
Background	Variant histone H2A which replaces conventional H2A in a subset of nucleosomes. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Required for checkpoint-mediated arrest of cell cycle progression in response to low doses of ionizing radiation and for efficient repair of DNA double strand breaks (DSBs) specifically when modified by C-terminal phosphorylation.
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
FCM	1:20-100
IHC-P	1:200-400
IHC-F	1:100-500
IF(IHC-P)	1:50-200
IF(IHC-F)	1:50-200
IF(ICC)	1:50-200