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Synthetic peptide corresponding to Human Histone H2A aa 1-15. This antibody was generated by immunizing rabbits with a mixture of synthetic peptides containing amino acid residues 1-15 (MSGRGKQGGKARAKA) and 81-96 (PRHLQLAIRNDEELNK) of human Histone H2A.
PRHLQLAIRNDEELNK (1-15), PRHLQLAIRNDEELNK (81-96)
Our Abpromise guarantee covers the use of ab13923 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 5 µg/ml.|
See Dai et al, 2008 for details on Nucleosome ELISA
|WB||Use a concentration of 2 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 16 kDa).|
|ChIP||Use at an assay dependent concentration. PubMed: 20711347|
ICC/IF image of ab13923 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab13923, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) ab150077) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Western blot analysis of Histone H2A in human PBMC cell lysate with ab13923.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast tissue (ab30090)labelling Histone H2A with ab13923 at 5µg/ml. Staining was enhanced by boiling tissue sections in 10mM sodium citrate buffer, pH6.0 for 10-20 minutes followed by cooling at room temperature for 20 minutes.
ab13923 staining Human normal skin (ab30166). Staining is localised to nuclear compartment.
Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
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