The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/5000. Detects a band of approximately 120 kDa (predicted molecular weight: 41 kDa).
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ab199507 - Rabbit monoclonal IgG (HRP), is suitable for use an as isotype control with this antibody.
Possible adhesion molecule with a role in early hematopoiesis by mediating the attachment of stem cells to the bone marrow extracellular matrix or directly to stromal cells. Could act as a scaffold for the attachment of lineage specific glycans, allowing stem cells to bind to lectins expressed by stromal cells or other marrow components. Presents carbohydrate ligands to selectins.
Selectively expressed on hematopoietic progenitor cells and the small vessel endothelium of a variety of tissues.
Belongs to the CD34 family.
On early hematopoietic progenitor cells.
Highly glycosylated. Phosphorylated on serine residues by PKC.
IHC image of CD34 staining in a section of formalin-fixed paraffin-embedded normal humrn umbilical cord tissue*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab195017 at 1/100 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
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.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Western blot - Anti-CD34 antibody [EP373Y] (HRP) (ab195017)
Anti-CD34 antibody [EP373Y] (HRP) (ab195017) at 1/5000 dilution + TF-1 (Human erythroleukaemic cell line) Whole Cell Lysate at 10 µg
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab195017 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.