Human Histone H2A.X Ready-To-Use IHC Kit

Cat. No.: IHC0308H
Sample Type: FFPE tissue
Size: 50T (including a control slide)
Storage and Stability: Please store components at the temperatures indicated on the individual tube labels. The kit is stable for 6 months from the date of receipt.

Background

Histone H2A.X (H2AX) is a member of the histone H2A family which is one of the four core histones making up the nucleosome core particle. In eukaryotes, DNA double strand breaks (DSBs) have been shown to trigger the phosphorylation of serine 139 at the carboxy terminus of histone H2AX resulting in gamma-H2AX. The phosphorylation of H2AX can be detected by Western blotting or immunofluorescence, revealing the frequency of DSBs. The phosphatidylinositol 3-kinases have been implicated in H2AX phosphorylation, but it is unclear if ATM is the primary H2AX kinase or if other members of the family such as DNA-PK and ATR contribute in a similar manner. Structurally, H2A.x contains 143 amino acid residues. Histone H2A.X is considered a basal histone, being synthesized in G1 as well as in S-phase, and its mRNA contains polyA addition motifs and a polyA tail along with the conserved stem-loop and U7 binding sequences involved in the processing and stability of replication type histone mRNAs. There are two forms of Histone H2A.X mRNA, one about 1600 bases long and contains polyA; the other about 575 bases long, lacking polyA. The short form behaves as a replication type histone mRNA, while the longer behaves as a basal type histone mRNA. Histone H2A.X maps to the 11q23.2-q23.3 region of the human chromosome. Histone H2A.x contributes to histone-formation and therefore the structure of DNA. Histone H2A variant H2A.x specifically regulates the interaction of MDC1 (mediator of DNA damage checkpoint protein 1), a DNA repair protein to the sites of DNA damage.

Synonyms

Materials Supplied

Number	Component	50T	Concentration	Storage
1	PBS Buffer (powder)	2L x 2	20x	RT
2	Antigen Retrieval Buffer	20ml	100x	2-8°C
3	Endogenous Peroxidase Blocking Buffer	3ml	RTU	2-8°C
4	Blocking Buffer	3ml	RTU	2-8°C
5	Primary Antibody (Human Histone H2A.X Recombinant Rabbit mAb)	6ml	RTU	2-8°C
6	Secondary Antibody (Goat Anti-Rabbit IgG H&L, HRP conjugated)	6ml	RTU	2-8°C
7	Chromogen Component A	0.3ml	RTU	-20°C
8	Chromogen Component B	0.3ml	RTU	-20°C
9	Counter Staining Reagent	5ml	RTU	RT
10	Differentiation Reagent	6ml	RTU	RT
11	Mounting Media	5ml	RTU	RT
12	Control slide (Human colon)	1 slide	RTU	RT
13	Datasheet	1 copy

Notes

1. The positive control slide provided in the kit allows you to be sure that the experimental set-up is working properly.
2. Do not allow slides to dry at any time during this procedure.
3. Please don't replace the matching reagents in this product with other manufacturers' products.
4. As DAB is a carcinogen, please take necessary precautions.
5. PBS (reagent 1) can be stored for one week at 4℃ after preparation; The antigen retrieval buffer (1×reagent 2) and the chromogenic agent (the mixture of reagents 7 and 8) should be prepared right before each assay.

As DAB is a carcinogen, please take necessary precautions.

Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.