Anti-Histone H3 (phospho S28) antibody [E191]

• Supplier: Abcam
• Catalog: ab32388
• Prices: $390.00
• Sizes: 100 µl
• Host: Rabbit
• Clonality: Monoclonal
• Isotype: IgG
• Clone: E191
• Applications: WB IHC-P ICC/IF ICC/IF FC IHC-F
• Species Reactivities: Mouse, Rat, Guinea Pig, Human
• Antigen: Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Histone H3 aa 1-100 (phospho S28)
• Description: Rabbit Monoclonal
• Gene: H3F3A
• Conjugate: Unconjugated

Product images

Dot blot performed using ab32388 at a dilution of 1/100. Lane 1 - Unmodified H3 peptide. Lane 2 - H3S28ph peptide. Lane 3 - H3.3S28ph peptide. Lane 4 - H3.3S31ph peptide. A HRP conjugated goat anti-rabbit (H+L) was used as the secondary antibody at a dilution of 1/2500. The exposure time was 3 minutes and the dilution and blocking buffer used were 5% NFDM/TBST.

All lanes : Anti-Histone H3 (phospho S28) antibody [E191] (ab32388) at 1/2000 dilutionLane 1 : NIH 3T3 cell lysate -untreatedLane 2 : NIH 3T3 cell lysate -treated with FBS + CalA.

IHC image of ab32388 staining Histone H3 in Human normal colon 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 ab32388, 0.1ug/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.*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

Overlay histogram showing HeLa cells stained with ab32388 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32388, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

Immunohistochemical analysis of Histone H3 (phospho S28) expression in paraffin embedded lymphoma tissue sections, using 1/100 ab32388. left: untreated sample; right: phosphatase treated sample (negative control).Immunohistochemical analysis of Histone H3 (phospho S28) expression in paraffin embedded lymphoma tissue sections, using 1/100 ab32388. left: untreated sample; right: phosphatase treated sample (negative control).

Product References

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