Purification of primary malignant B-cells and immunoblot analysis of bcl-2 family proteins.
The ability to isolate relatively pure populations of primary normal and malignant lymphocytes has brought studies of lymphoid malignancies to the forefront of cancer research. Apoptosis (programmed cell death) plays a key role in controlling normal B-cell numbers, and resistance to apoptosis contributes to lymphomagenesis and reduces the effectiveness of chemotherapy and radiotherapy. Multiple Bcl-2 family proteins orchestrate key life and death decisions in lymphocytes, and the prototypical family member, Bcl-2, is activated by reciprocal translocation in human lymphoma. Here, we describe an immunomagnetic method to isolate and purify malignant B-cells suitable for in vitro analyses from lymph node biopsies. Methods to analyze the expression of Bcl-2 family proteins by immunoblotting also are described.
Cancer Sciences Division, University of Southampton, UK.
Claire
C
Dallman
Graham
G
Packham
D001402:B-Lymphocytes / Q000473:pathology*; D001706:Biopsy; D006801:Humans; D015151:Immunoblotting; D018189:Immunomagnetic Separation / Q000379:methods*; D008198:Lymph Nodes / Q000473:pathology; D016393:Lymphoma, B-Cell / Q000473:pathology*; D008565:Membrane Proteins / Q000032:analysis; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D009363:Neoplasm Proteins / Q000032:analysis; D011518:Proto-Oncogene Proteins / Q000032:analysis; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000032:analysis*; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein
C494811:BCL2L1 protein, human; C090121:BNIP3 protein, human; D008565:Membrane Proteins; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D009363:Neoplasm Proteins; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein