Histone H2A.X (Phospho-S139) Rabbit Polyclonal Antibody

MSDS

Histone H2A.X (Phospho-S139) Rabbit Polyclonal Antibody

Key features and details

Rabbit polyclonal antibody to Histone H2A.X (Phospho-S139)

Target: Histone H2A.X (Phospho-S139)
Source/Host: Rabbit
Reactivity: Human, Mouse, Rat
Clonality: Polyclonal
Applications: WB, IHC, IF/ICC, FC
Conjugation: Unconjugated
Storage: at-20°C

Brand:

CAT.NO. : APA10259

US$ Please choose

Size:

50μL 100μL

Trail, Bulk size or Custom requests Please contact us

Product Details

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

To ensure optimal assay performance, AREX recommends conducting reagent titration tailored to each testing system for optimal detection results.

WB	1:500 - 1:1000
IHC	1:50 - 1:100
IF/ICC	1:50 - 1:200
FC	1:100 - 1:200

*Results are sample-specific. Please refer to your local assay conditions and test parameters for reference.

Overview

Description	Rabbit polyclonal antibody to Histone H2A.X (Phospho-S139)
Specificity	Recognizes endogenous levels of Histone H2A.X protein only when phosphorylated at S139.
Antibody Type	Primary antibody
Imnunogen	KLH-conjugated synthetic phosphopeptide corresponding to residues surrounding S139 of human Histone H2A.X protein. The exact sequence is proprietary.
Purification	The antibody was purified by immunogen affinity chromatography.
Molecular Weight	Predicted: 15 kD; Observed: 20 kD
Form/Buffer	Liquid in 0.42% Potassium phosphate, 0.87% Sodium chloride, pH 7.3, 30% glycerol, and 0.01% sodium azide.
Alternative Names	H2AX; Histone H2AX; H2a/x; Histone H2A.X
Gene Symbol	H2AFX
Entrez Gene	3014(Human); 15270(Mouse)
SwissProt	P16104(Human); P27661(Mouse)

*AREX continuously optimizes our products. Webpage content may not reflect the latest updates. For inquiries, please contact info@arexbio.com or your local distributor.
*Clone Number, Reactivity, Source/Host and Clonality can be found in the product name and Key Features section above.

Data

Western blot analysis of Histone H2A.X (Phospho-S139) expression in HEK293T (A) whole cell lysates. (Predicted band size: 15 kD; Observed band size: 20 kD)

Immunohistochemical analysis of Histone H2A.X (Phospho-S139) staining in human breast cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of Histone H2A.X (Phospho-S139) staining in C6 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 °C in a hidified chamber. Cells were washed with PBST and incubated with a AREX® Fluor 488 -conjugated secondary antibody (green) in PBS at room temperature in the dark. Phalloidin - AREX® Fluor 594 was used to stain Actin filaments (red). DAPI was used to stain the cell nuclei (blue).

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Note

For Research Use Only. Not for use in diagnostic procedures.

FAQs

What are the main types of research antibodies and how do they differ?

Research antibodies are mainly divided into monoclonal antibodies and polyclonal antibodies. Monoclonal antibodies typically offer higher specificity and better batch-to-batch consistency, while polyclonal antibodies often provide stronger affinity but may show more variation between batches. The choice depends on your specific experimental needs.

How can I tell if a research antibody is suitable for my experiment?

It is recommended to carefully review the product datasheet for validated applications, species reactivity, recommended dilutions, and published references. For new antibodies, performing a small-scale validation with positive control samples is usually helpful.

Can improper storage of research antibodies affect experimental results?

Yes. Antibodies are sensitive to temperature, repeated freeze-thaw cycles, and contamination. Improper storage may lead to reduced activity, increased background, or weaker signals. It is best to follow the storage instructions provided in the product datasheet.

Why doesn’t the recommended dilution in the datasheet work well in my experiment?

The recommended dilution is based on the supplier’s test conditions. Factors such as sample type, fixation method, and detection system in your lab can influence the optimal working concentration. Performing a dilution series optimization in your own system is often necessary.

What precautions should I take when using a newly purchased research antibody for the first time?

It is advisable to briefly centrifuge the antibody (especially concentrated or lyophilized ones), then perform a small-scale pilot experiment using the recommended conditions. Recording the batch number and usage date is also helpful for future tracking.

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