Clone: VIT200 and MEM18
Label: FITC,PE
Background: The CD45 molecule is typically expressed at high levels on all hematopoietic cells. CD45 is a major component of the glycocalix of these cells and can be expressed in different isoforms. Antibody VIT200 recognizes a pan CD45 epitope, which is expressed on all hematopoietic cells. CD14 is a GPI-anchored molecule expressed by virtually all human monocytes and macrophages and – to a lesser degree - granulocytes. CD14 together with Toll-like receptor 4 and MD-2 forms the LPS-receptor complex that recognizes and signals the presence of LPS. While CD14 has no signaling structure its main role seems to be the binding of LPS. The VIT200/MEM18 COMBI-REAGENT-Gate Control permits the identification and enumeration of human leukocytes using flow cytometry.Results must be interpreted by a certified professional before final interpretation. Analyses performed with this antibody should be paralleled by positive and negative controls. If unexpected results are obtained which cannot be attributed to differences in laboratory procedures, please contact us.
Specificity: The CD45 mAb (clone VIT200) recognizes a pan CD45 epitope. The CD14 mAb (clone MEM18) recognizes surface CD14 on human monocytes and macrophages as well as neutrophils. The sensitivity of CD45/CD14 mAb is determined by staining well-defined blood samples from representative donors with serial-fold mAb dilutions to obtain a titration curve that allows relating the mAb concentration to the percentage of stained cells and geometric MFI (mean fluorescence intensity). For this purpose, a mAb-concentration range is selected to include both the saturation point (i.e. the mAb dilution expected to bind all epitopes on the target cell) and the detection threshold (i.e. the mAb dilution expected to represent the least amount of mAb needed to detect an identical percentage of cells). In practice, 50 µl of leukocytes containing 10^7 cells/ml are stained with 20 µl mAb of various dilutions to obtain a titration curve and to identify the saturation point and detection threshold. The final concentration of the product is then adjusted to be at least 3-fold above the detection threshold. In addition and to control lot-to-lot variation, the given lot is compared and adjusted to fluorescence standards with defined intensity.
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Caution: For professional users only. This reagent contains sodium azide. To avoid the development of hazardous conditions, reagents containing azide should be diluted in running water prior to be discarded. Similar to the work with other biological products, proper handling procedures are recommended.
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