Overlay histogram showing Jurkat cells stained with ab8536 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab8536, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in Jurkat cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.

Anti-CD34 antibody [QBEND-10] (ab8536) at 5 µg/ml + Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate at 10 µg

Secondary
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 40 kDa
Observed band size: 40 kDa
Additional bands at: 100 kDa (possible glycosylated form)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labelling CD34 with ab8536 at 1µg/ml (1:1000). Blood vessels show positive staining.

Detection system polymeric HRP. HIER with Pressure cooker pH6.

ab8536 staining CD34 in human kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/2000 in blocking buffer) for 16 hours at 21°C. A Biotin-conjugated goat anti-mouse IgG polyclonal (1/300) was used as the secondary antibody.

See Abreview

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labelling CD34 with ab8536 at 1µg/ml (1:1000). Blood vessels show positive staining.

Detection system polymeric HRP. HIER with Pressure cooker pH6.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labeling CD34 with ab8536 at 1ug/ml.
Detection system polymeric HRP.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human umbilical cord labeling CD34 with ab8536 at 1/1000 dilution. Tissue was fixed in formaldehyde and permeabilized with PBS. Heat mediated antigen retrieval was performed using a citrate pH 6 buffer. Tissue was blocked with 5% serum for 1 hour at room temperature. A biotinylated polyclonal goat anti-mouse secondary antibody was used at 1/100 dilution.

See Abreview

ab8536 at 1/500 staining human umbilical cord tissue sections by IHC-P. The tissue was paraformaldehyde fixed and blocked with horse serum before incubation with the antibody for 16 hours. A biotinylated goat anti-mouse IgG was used as the secondary.

See Abreview