Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labeling Hemoglobin subunit alpha antibody with purified ab92492 at 1/500 dilution (1.4 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. ab97051, a HRP-conjugated goat anti-rabbit IgG H&L was used as the secondary antibody at 1/500 dilution. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92492).

Clone EPR3608 (ab174536) has been successfully conjugated by Abcam. This image was generated using Anti-Hemoglobin subunit alpha antibody [EPR3608] (Alexa Fluor® 647). Please refer to ab215919 for protocol details.

IHC image of Hemoglobin subunit alpha staining in a section of formalin-fixed paraffin-embedded normal human placenta*.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Dako Pascal pressure cooker using the standard factory-set regime. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature.  The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab215919 at 1/100 dilution (shown in red) and counterstained using ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount^®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebral cortex tissue labeling Hemoglobin subunit alpha antibody with purified ab92492 at 1/500 dilution (1.4 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. ab97051, a HRP-conjugated goat anti-rabbit IgG H&L was used as the secondary antibody at 1/500 dilution. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92492).

Immunohistochemical analysis of paraffin embedded human placenta tissue labeling hemoglobin subunit alpha with unpurified ab92492 at 0.051 µg/ml (1:40000 dilution).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92492).

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin embedded human placenta tissue labeling hemoglobin subunit alpha with unpurified ab92492 at 0.051 µg/ml (1:40000 dilution).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92492).

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

This IHC data was generated using the same anti-hemoglobin subunit alpha antibody clone, EPR3608, in a different buffer formulation (cat# ab924920).

Immunohistochemical analysis of paraffin embedded human stomach cancer tissue labeling hemoglobin subunit alpha with unpurified ab92492 at 0.051 µg/ml (1:40000 dilution).

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

This IHC data was generated using the same anti-hemoglobin subunit alpha antibody clone, EPR3608, in a different buffer formulation (cat# ab924920).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human placenta tissue labeling Hemoglobin subunit alpha antibody with purified ab92492 at 1/500 dilution (1.4 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. ab97051, a HRP-conjugated goat anti-rabbit IgG H&L was used as the secondary antibody at 1/500 dilution. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.