Anti-CD34 抗体 [MEC 14.7] (ab8158)

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.

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

Human aortic valve: immunofluorescence labelling shows telocytes.

Frozen sections were sliced into a thickness of 6 μM, and then were post‐fixed with 4% paraformaldehyde dissolved in 0.1 M phosphate buffer (pH = 7.4) for at least 15 min. After washed with phosphate buffer for three times, sections were immersed in 10% goat serum for 1 hr. After that, sections were incubated overnight at 4°C with rat monoclonal anti‐CD34 (ab8158; Abcam, Cambridge, UK) and either rabbit monoclonal to Vimentin (ab92547; Abcam), rabbit monoclonal to PDGF Receptor‐beta (ab32570, 1:100; Abcam) or rabbit polyclonal anti‐C‐Kit (ab5506, 1:100; Abcam), both were with the dilution of 1:100 in phosphate buffer and permeabilized by 0.25% Triton X‐100 at the same time. On the second day, the sections were incubated to goat anti‐rabbit labelled with rhodamine secondary antibodies and goat anti‐rat labelled with FITC diluted 1:200 in phosphate buffer for 1 hr. The sections were stained with 4′,6‐diamidino‐2‐phenylindole (DAPI). 

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.

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ab8158 staining CD34 in mouse lung endothelial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde and blocked with 1% BSA for 30 minutes at 4°C. Samples were incubated with primary antibody (1/100) for 16 hours at 4°C. An undiluted Alexa Fluor® 555-conjugated goat anti-rat IgG polyclonal 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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Ab8158 at a dilution of 1/50 staining CD34 from mouse lung cells by Immunocytochemistry. The antibody was incubated with the sample for 1 hour and then detected using an Alexa-Fluor 488® goat anti-rat antibody. The staining for CD34 is shown in green. The samples were also stained for Smooth Muscle Actin (red stain).

This image is courtesy of an Abreview submitted on 8 February 2006.

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