WB result of Histone H2A.X Recombinant Rabbit mAb
Primary antibody: Histone H2A.X Recombinant Rabbit mAb at 1/5000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: MCF7 whole cell lysate 20 µg
Lane 3: HepG2 whole cell lysate 20 µg
Lane 4: A431 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
Histone H2A.X Recombinant Rabbit mAb (S-1772-179)
Histone H2A.X Recombinant Rabbit mAb (S-1772-179)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Histone H2A.X |
| Synonyms | Histone H2AX; H2a/x; Histone H2A.X; H2AFX; H2AX |
| Immunogen | Synthetic Peptide |
| Location | Nucleus |
| Accession | P16104 |
| Clone Number | S-1772-179 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ChIP |
| Reactivity | Hu, Ms, Rt |
| Positive Sample | HeLa, MCF7, HepG2, A431, NIH/3T3 |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:5000 | Hu, Ms |
| IHC-P | 1:2000 | Hu, Ms, Rt |
| ChIP | 1:20-1:50 | Hu |
Background
Histone H2A.X is a histone variant found in almost all eukaryotes that plays a crucial role in maintaining genome stability. It is highly conserved across species and has a unique C-terminal tail containing an SQ motif, which is phosphorylated by kinases like ATM and ATR in response to DNA double-strand breaks (DSBs) to form γ-H2A.X. This phosphorylation event is a very early and highly conserved response to DNA damage, occurring within minutes in mammals and extending over large chromatin regions around the damage site. γ-H2A.X acts as a major signal to recruit DNA repair factors to the damaged site, facilitating efficient repair and preserving genomic integrity. Deficiencies in H2A.X or its phosphorylation can lead to genomic instability, increased sensitivity to DNA-damaging agents, and a higher risk of tumor formation. Additionally, H2A.X is involved in various other cellular processes, including the regulation of gene expression, immune response, and the establishment of cell fate.
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Picture
Western Blot
WB result of Histone H2A.X Recombinant Rabbit mAb
Primary antibody: Histone H2A.X Recombinant Rabbit mAb at 1/5000 dilution
Lane 1: NIH/3T3 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human testis. Anti-Histone H2A.X antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti-Histone H2A.X antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse liver. Anti-Histone H2A.X antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat cerebral cortex. Anti-Histone H2A.X antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
ChIP
Chromatin immunoprecipitation (ChIP) was
performed on HeLa cells cross - linked with 1% formaldehyde
for 10 min, then chromatin was fragmented by sonication.
Parallel reactions used Histone H2A.X Recombinant Rabbit
mAb (S-1772-179) and Rabbit mAb IgG Isotype Control
(SDT-R173) at 1:50 for immunoprecipitation.
Post - immunoprecipitation, both samples
were washed, eluted and cross-links
reversed. Purified DNA was analyzed by qPCR.
qPCR (%input: immunoprecipitated DNA/input DNA)
showed the enrichment of RPL30, GAPDH, AFM, MYOD1, SAT-α and SAT-2 in Histone H2A.X Recombinant Rabbit mAb
(S-1772-179)-immunoprecipitated sample.
