Detection of Human, Mouse, and Rat Histone H2AY by Western Blot.
Western blot shows lysates of MCF‑7 human breast cancer cell line, Hepa 1‑6 mouse hepatoma cell line, and Rat‑2 rat embryonic fibroblast cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse/Rat Histone H2AY Monoclonal Antibody (Catalog # MAB8318) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Histone H2AY at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Histone H2AY in HeLa Human Cell Line.
Histone H2AY was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human/Mouse/Rat Histone H2AY Monoclonal Antibody (Catalog # MAB8318) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (yellow, upper panel; Catalog # NL007) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Histone H2AY
Histone H2AY, also known as macro H2A1 (MH2A1) and H2AFY, is a 40 kDa variant of the histone family, basic nuclear proteins responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Ubiquitously expressed, H2AY replaces conventional H2A histones in a subset of nucleosomes, where it represses transcription and participates in stable X chromosome inactivation. H2AY expression is also enriched in senescence-associated heterochromatin. Alternative splicing creates multiple transcript variants, encoding three distinct isoforms. H2AY expressed in blood has been identified as a potential biomarker associated with the activity and therapeutic response of Huntington disease, an autosomal dominant neurodegenerative disorder. Over amino acids 230-348, human H2AY shares 98% identity with mouse and rat H2AY.
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