Anti-Histone H2A (symmetric di methyl R3)抗体

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A (symmetric di methyl R3) antibody (AB22397)

ICC/IF image of ab22397 stained human MCF7 cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab22397, 5μg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, HEK 293 and HepG2 cells.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A (symmetric di methyl R3) antibody (AB22397)

IHC image of ab22397 staining Histone H2A (symmetric di methyl R3) in human kidney formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab22397, 5μg/ml, for 15 mins 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. No primary antibody was used in the negative control (shown on the inset).

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• WB

Lab

Western blot - Anti-Histone H2A (symmetric di methyl R3) antibody (AB22397)

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab22397 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-Histone H2A (symmetric di methyl R3) antibody (ab22397) at 1 µg/mL

Lane 1:

Calf Thymus Histone at 0.5 µg

Lane 2:

HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg

Lane 3:

Histone H2A Recombinant at 0.1 µg

Lane 4:

Histone H3.1 Recombinant (negative control) at 0.1 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

Predicted band size: 14 kDa

Observed band size: 17 kDa

true

Exposure time: 2min

• ELISA

Unknown

ELISA - Anti-Histone H2A (symmetric di methyl R3) antibody (AB22397)

This antibody gave a positive result in ELISA against the immunizing peptide ab22399 (blue line). Histone H2A (unmodified) peptide ab13186 (pink line) gave a negative result, This indicates that it is specific for the Histone H2A (symmetric di methyl R3) peptide.