CD34 Monoclonal Antibody (4H11), Super Bright™ 600, eBioscience™, Invitrogen™

Description

Description: The 4H11 monoclonal antibody reacts with human CD34, also known as mucosialin. CD34 belongs to a protein family which also includes endoglycan and podocalyxin. Members of this family are single pass transmembrane proteins with a heavily glycosylated extracellular and N-terminal mucin domain. CD34 was first identified as an antigen expressed on hematopoietic progenitors, and has since been extensively used as a marker to isolate cells capable of hematopoietic cell engraftment. In spite of this, the function of CD34 remains unresolved. In addition to expression on hematopoietic progenitors, CD34 is expressed on some populations of mesenchymal stem cells, tumor cell lines, and by vascular endothelia in the adult. Epitopes of CD34 have been assigned to three classes (class I, II or III) based on their differential sensitivity to enzymatic cleavage by neuraminidase, chymopapain, or O-glycoprotease. According to this analysis, the 4H11 antibody belongs to class III, indicating that it reacts with a protein epitope. Applications Reported: This 4H11 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 4H11 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 μL (0.25 μg) per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL.

Cell number should be determined empirically but can range from 10⁵ to 10⁸ cells/test. Super Bright 600 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 600 nm. We recommend using a 610/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product No. SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 2-8°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorochrome performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 405 nm; Emission: 600 nm; Laser: Violet Laser Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company. CD34 is a highly glycosylated, monomeric, 111-115 kDa surface protein that is present on many stem cell populations. CD34 is a stem cell marker although its expression on human hematopoietic stem cells is reversible. CD34 may serve as a surface receptor that undergoes receptor-mediated endocytosis and regulates adhesion, differentiation, and proliferation of hematopoietic stem cells and other progenitors. CD34 expression is likely to represent a specific state of hematopoietic development that may have altered adhering properties with expanding and differentiating capabilities in both in vitro and in vivo conditions. CD34 is possibly an adhesion molecule with a putative role for mediating the attachment of stem cells to the bone marrow extracellular matrix or directly to stromal cells. Further, CD34 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. CD34 is thought to have a role in presenting carbohydrate ligands to selectins. The intracellular chain of the CD34 antigen is a site of phosphorylation by activated protein kinase C suggesting a putative role in signal transduction. Diseases associated with CD34 dysfunction include dermatofibrosarcoma and neurofibroma.