Anti-HLA-DR antibody [DA6.147] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [DA6.147] - BSA and Azide free (AB244584)

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab238469).

IHC image of HLA-DR staining in a section of formalin-fixed paraffin-embedded normal human breast* performed on a Leica BOND<™> system using the standard Protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab238469, 0.5μg/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. The inset secondary-only control image is taken from an identical assay without primary antibody.

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA-DR antibody [DA6.147] - BSA and Azide free (AB244584)

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab238469).

IHC image of HLA-DR staining in a section of formalin-fixed paraffin-embedded normal human tonsil* performed on a Leica BOND<™> system using the standard Protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab238469, 0.5μg/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. The inset secondary-only control image is taken from an identical assay without primary antibody.

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.

• WB

Lab

Western blot - Anti-HLA-DR antibody [DA6.147] - BSA and Azide free (AB244584)

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab238469).

This blot was produced using a 4-12% Bis-tris under the MES buffer system. The gel was run at 200V for m5 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour using 3% milk before ab238469 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at a 1ug/ml concentration and 1/20000 dilution respectively. Antibody binding was detected using Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-HLA-DR antibody [DA6.147] (<a href='/en-us/products/primary-antibodies/hla-dr-antibody-da6147-ab238469'>ab238469</a>) at 1 µg/mL

Lane 1:

Raji whole cell lysate at 20 µg

Lane 2:

Ramos whole cell lysate at 20 µg

Predicted band size: 29 kDa

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