IHC image of CD34 staining in a formalin fixed paraffin embedded mouse lung tissue section, performed on a Leica Bond™ system using the standard protocol B. 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 ab8158, 1 µg/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.

Anti-CD34 antibody [MEC 14.7] (ab8158) at 1 µg/ml + Mouse Lung Membrane Tissue Lysate (ab171830) at 50 µg

Secondary
Goat Anti-Rat IgG H&L (HRP) (ab97057) at 1/20000 dilution

Performed under reducing conditions.

Predicted band size: 41 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?


Exposure time: 2 minutes

The main band is as expected at 80 kDa since the target is heavily glycosylated and phosphorylated.

IHC image of CD34 staining in a formalin fixed paraffin embedded mouse brain tissue section, performed on a Leica Bond™ system using the standard protocol B. 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 ab8158, 10 µg/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.

Immunohistochemical analysis of murine lung tissue, staining CD34 with ab8158.

Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at room temperature; antigen retrieval was by heat mediation in citrate buffer (pH 6). Samples were incubated with primary antibody (1/100 in diluent) for 16 hours at 4°C. An undiluted HRP-conjugated horse anti-rat polyclonal IgG was used as the secondary antibody.

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ab8158 staining CD34 in mouse mammary gland tissue sections by Immunohistochemistry (IHC-P - paraffin-embedded sections). Tissue was fixed with methacarnoy and blocked with 4% BSA for 60 minutes; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100) for 16 hours. A undiluted HRP-conjugated goat anti-rat IgG polyclonal was used as the secondary antibody.

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ab8158 staining mouse brain tissue sections by IHC-P.  Tissue sections were paraformaldehyde fixed and blocked with 0.5% Perkin-Elmer TNB Blocking Buffer for 30 minutes at 25°C.  The primary antibody was diluted 1/100 and incubated for 18 hours at 4°C.  An biotin conjugated goat anti-rat was used as the secondary

Photomicrograph demonstrates CD34 in red and collagen type IV (ab19808) in blue in normal, adult brain vessels. Tissue was perfusion-fixed and cut into 15µm slide-mounted cryostat sections (i.e., lightly fixed, but not paraffin embedded).

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