Anti-CD34 antibody [EP373Y]

14 product images

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  Flow Cytometry (Intracellular) - Anti-CD34 antibody [EP373Y] (ab81289)

  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.
  

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  This image is courtesy of a customer review submitted by Rudolf Jung.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD34 antibody [EP373Y] (ab81289)

  Paraformaldehyde-fixed, paraffin-embedded mouse kidney tissue stained for CD34 using ab81289 at 1/200 dilution in immunohistochemical analysis.

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  This image is courtesy of an anonymous customer review.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD34 antibody [EP373Y] (ab81289)

  Paraformaldehyde-fixed, paraffin-embedded mouse prostate tissue stained for CD34 using ab81289 at 1/150 dilution in immunohistochemical analysis.

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  Immunoprecipitation - Anti-CD34 antibody [EP373Y] (ab81289)

  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

  All lanes: Immunoprecipitation - Anti-CD34 antibody [EP373Y] (ab81289)

  Predicted band size: 40 kDa

  Observed band size: 120 kDa

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  Immunohistochemistry (Frozen sections) - Anti-CD34 antibody [EP373Y] (ab81289)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat kidney tissue labeling CD34 with ab81289 at 1/50 (11.04 μg/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on rat kidney. is observed. The nuclear counterstain was DAPI (Blue).
  

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

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

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  Immunohistochemistry (Frozen sections) - Anti-CD34 antibody [EP373Y] (ab81289)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat lung tissue labeling CD34 with ab81289 at 1/50 (11.04 μg/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on rat lung. is observed. The nuclear counterstain was DAPI (Blue).
  

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

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

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  Immunocytochemistry/ Immunofluorescence - Anti-CD34 antibody [EP373Y] (ab81289)

  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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counter stain.
  

  Control: Secondary antibody only, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 488-conjugated goat anti-mouse IgG (1/500).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD34 antibody [EP373Y] (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) (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.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD34 antibody [EP373Y] (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) (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.

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  OI-RD Scanning - Anti-CD34 antibody [EP373Y] (ab81289)

  We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
  Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD34 antibody [EP373Y] (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.

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  Image from Földes G et al. PLoS One. 2010 May 5;5(5):e10501 Fig 3.

  Immunocytochemistry/ Immunofluorescence - Anti-CD34 antibody [EP373Y] (ab81289)

  CD34 Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-CD34 antibody

  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.

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  Flow Cytometry (Intracellular) - Anti-CD34 antibody [EP373Y] (ab81289)

  Flow cytometry overlay histogram showing left HUVEC positive cells and right negative HL60 stained with ab81289 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 mins. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab81289) (1x 10⁶ in 100μl at 0.2μg/ml (1/11550 dilution)) for 30 mins at 22°C.

  The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 dilution for 30 mins at 22°C

  Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

  Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

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  Western blot - Anti-CD34 antibody [EP373Y] (ab81289)

  CD34 Western blot staining using rabbit Anti-CD34 antibody

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  Negative control: Daudi, HL-60.

  Low expression: HUVEC, Neuro-2a.

  All lanes: Western blot - Anti-CD34 antibody [EP373Y] (ab81289) at 1/1000 dilution

  Lane 1: KG-1a (human bone marrow myeloblast) whole cell lysate at 20 µg

  Lane 2: TF-1 (human erythroleukemia erythroblast) whole cell lysate at 20 µg

  Lane 3: Daudi (human Burkitts lymphoma lymphoblast) whole cell lysate at 20 µg

  Lane 4: HL-60 (Human acute promyelocytic leukemia promyeloblast) whole cell lysate at 20 µg

  Lane 5: HUVEC (Human umbilical vein endothelial cell) whole cell lysate at 20 µg

  Lane 6: bEnd.3 (mouse brain endothelioma) whole cell lysate at 20 µg

  Lane 7: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

  Lane 8: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 80-120 kDa

  Exposure time: 60s