Recombinant Anti-CD34 antibody [EP373Y] - BSA and Azide free (ab198395)

Intracellular Flow Cytometry analysis of TF-1 (human erythroleukemia) cells labeling CD34 with purified ab81289 at 1/50 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. 

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81289). 

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81289).

See Abreview

This data was developed using ab81289, the same antibody clone in a different buffer formulation.

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution.<\p>

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

This data was developed using ab81289, the same antibody clone in a different buffer formulation.

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution.<\p>

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81289).

See Abreview

CD34 was immunoprecipitated from TF-1 (Human bone marrow erythroleukemia cells) cells using purified ab81289 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab81289. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as secondary antibody at 1/1000 dilution. Blocking and dilution buffer and concentration: 5% NFDM/TBST

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81289).

Clone EP373Y (ab198395) has been successfully conjugated by Abcam. This image was generated using Anti-CD34 antibody [EP373Y] (Alexa Fluor^® 488). Please refer to ab195013 for protocol details. 

Overlay histogram showing HUVEC cells stained with ab195013 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab195013, 1/500 dilution) for 30 min at 22°C. 

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor^® 488 (ab199091) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. 

Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. 

This antibody gave a positive signal in HUVEC cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions. 

Clone EP373Y (ab198395) has been successfully conjugated by Abcam. This image was generated using Anti-CD34 antibody [EP373Y] (PE). Please refer to ab223930 for protocol details. 

Overlay histogram showing HUVEC cells stained with ab223930 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab223930, 1/500 dilution) for 30 min at 22°C. 

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Phycoerythrin (ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. 

Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter. 

Immunocytochemistry/Immunofluorescence analysis of HUVEC (Human umbilical vein endothelial cell line) cells labelling CD34 with purified ab81289 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (ab150077) (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counter stain.

Control: Secondary antibody only, ab150120, an Alexa Fluor^® 488-conjugated goat anti-mouse IgG (1/500).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81289).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling CD34 with purified ab81289 at 1/2500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody at 1/500. Negative control using PBS instead of primary antibody. Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81289).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling CD34 with purified ab81289 at 1/2500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody at 1/500. Negative control using PBS instead of primary antibody. Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81289).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human angiosarcoma labeling CD34 with unpurified ab81289 at 1/100-1/250.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81289).

Immunocytochemistry/Immunofluorescence analysis of human embryonic stem cell-derived endothelial cells labeling CD34 with unpurified ab81289 at 1/200. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.2% aTriton X-100. An Alexa Fluor^® 647-conjugated secondary antibody was used at a 1/400 dilution. DAPI (blue) was used as the nuclear counter stain.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81289).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Normal kidney vessels tissue labeling CD34 with unpurified ab81289 at 1/100-1/250.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81289).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Normal uterus vessels tissue labeling CD34 with unpurified ab81289 at 1/100-1/250.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81289).

IHC image of CD34 staining in Mouse normal brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM 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 unpurified ab81289, 1/250 dilution, for 15 mins at room temperature. A Goat anti-Rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81289).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81289).

This IHC data was generated using the same anti-CD34 antibody clone, EP373Y, in a different buffer formulation (cat# ab81289).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Normal tonsil vessels tissue labeling CD34 with unpurified ab81289 at 1/100-1/250.