RosetteSep™ HLA B Cell Enrichment Cocktail

Procedures and instruction manuals:

•  PRODUCT INFORMATION SHEET - 15064HLA, Lot# 11J41202 or greater:29671-CE_Marked-PIS_2_0_0.pdf
•  PRODUCT INFORMATION SHEET - 15064HLA, Lot# 11G40696 or less:29058-PIS_1_0_0.pdf
•  PRODUCT INFORMATION SHEET - 15084HLA, Lot# 11J41202 or greater:29671-CE_Marked-PIS_2_0_0.pdf
•  PRODUCT INFORMATION SHEET - 15084HLA, Lot# 11G40696 or less:29058-PIS_1_0_0.pdf

Educational resources:

•  WEBINAR: Cell Separation Solutions for HLA and Chimerism Analysis
•  VIDEO: RosetteSep® Introduction
•  VIDEO: RosetteSep® for HLA Analysis
•  VIDEO: RosetteSep®: A Guide to Isolating NK Cells Directly from Whole Blood
•  BROCHURE: One-Step Cell Enrichment Directly From Whole Blood
•  BROCHURE: Cell Separation Products for HLA Analysis
•  BROCHURE: Ready Sep Go – Cell Separation In As Little As 25 Minutes
•  BROCHURE: Fast and Easy Isolation of B Cells
•  WALLCHART: Frequencies of Cell Types in Human Peripheral Blood Wallchart

MSDS:

•  15024HC.1-MSDS.pdf

FAQS:

• 
  Q. WHAT IS ROSETTESEP^®?
  
  A. RosetteSep^® is a rapid cell separation procedure for the isolation of purified cells directly from whole blood without columns or magnets.
• 
  Q. HOW DOES ROSETTESEP^® WORK?
  
  A. The antibody cocktail crosslinks unwanted cells to red blood cells (RBCs), forming rosettes. The unwanted cells then pellet with the free RBCs when centrifuged over a buoyant density medium.
• 
  Q. WHICH CELL SAMPLES CAN ROSETTESEP^® BE USED WITH?
  
  A. RosetteSep^® can be used with leukapheres, bone marrow samples or buffy coat, as long as: the concentration of cells does not exceed 5 x 10⁷ per mL (can dilute if necessary); and there are at least 20 RBCs for every nucleated cell (RBCs can be added if necessary).  For a full procedure click here.
• 
  Q. CAN ROSETTESEP^® BE USED WITH PREVIOUSLY FROZEN OR CULTURED CELLS?
  
  A. Yes. Cells should be re-suspended at 2 – 5 x 10⁷ cells / mL in PBS + 2% FBS. Fresh whole blood should be added at 250 µL per mL of sample, as a source of red cells.
• 
  Q. WHICH ANTICOAGULANT SHOULD BE USED WITH ROSETTESEP^®?
  
  A. Peripheral blood should be collected in heparinized Vaccutainers. Cord blood should be collected in ACD.
• 
  Q. SHOULD THE ANTICOAGULANT BE WASHED OFF BEFORE USING ROSETTESEP^®?
  
  A. No, the antibody cocktail can be added directly to the sample.
• 
  Q. WHICH FACTORS AFFECT CELL RECOVERY?
  
  A. The temperature of the reagents can affect cell recovery. All reagents should be at room temperature (sample, Ficoll^TM, PBS, centrifuge) before performing the isolations. Layering can also affect recovery so be sure to carefully layer the sample to avoid mixing with the Ficoll^TM as much as possible. Be sure to collect the entire enriched culture without disturbing the RBC pellet. A small amount of Ficoll^TM can be collected without worry.
• 
  Q. CAN ROSETTESEP^® BE USED TO ENRICH PROGENITORS FROM CORD BLOOD?
  
  A. Yes. Sometimes cord blood contains immature nucleated red cells that have a lower density than mature RBCs. These immature red cells do not pellet over Ficoll^TM, which can lead to a higher RBC contamination than peripheral blood separations.
• 
  Q. DOES ROSETTESEP^® WORK WITH MOUSE CELLS?
  
  A. No, but we have developed EasySep^®, a magnetic-based separation for the isolation of mouse cells.

FACS Histogram Results with RosetteSep® HLA B Cell Enrichment Kit

RosetteSep® Procedure