CD68, PE-eFluor 610, clone: eBioY1/82A (Y1/82A), eBioscience™

Description

CD68 (Macrosialin) is a 110kDa integral membrane glycoprotein predominantly expressed on the intracellular lysomsomes of monocytes and macrophages and to a lesser extent by dendritic cells and peripheral blood granulocytes. Elevated expression of CD68 has been demonstrated on CD34+ cells in various human malignancies, including several Acute Myeloid Leukemia studies.

Description: The eBioY1/82 antibody reacts with CD68 which belongs to the sialomucin family and is closely related to the family of lysosomal-associated membrane proteins (lamps) and scavenger receptor. CD68 is predominantly an intracellular protein, found mainly in the late endosomal compartment but can be detected in smaller amounts on the surface of cells of mainly myeloid derived cells; monocytes and macrophages, dendritic cells (and in some langerhan cells), neutrophils, basophils mast cells, myeloid progenitor cells, and a subset of CD34+ hemopoietic bone marrow progenitor cells. The function has not been fully elucidated but based on it's homolgy and structure suggests a role in antigen processing or presentation and protection of the lysosomal membrane from hydrolytic enzymes. Reports have also seen expression in activated T cells, and about 40% of peripheral blood B-lymphocytes and 50% of all B-ALL.Staining with eBioY1/82A shows surface staining on B cells and a minor percentage of monocytes while intracellular staining results in about 50% of B cells and most monocytes.Applications Reported: This eBioY1/82A (Y1/82A) antibody has been reported for use in intracellular staining followed by flow cytometric analysis.Applications Tested: This eBioY1/82A (Y1/82A) antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of normal human peripheral blood cells using the Intracellular Fixation & Permeabilization Buffer Set (cat. 88-8824) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resources Tab online. 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 10^5 to 10^8 cells/test. PE-eFluor™ 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red™). Please make sure that your instrument is capable of detecting this fluorochome.Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please 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 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.Filtration: 0.2μm post-manufacturing filtered.