Human PBMCs stained with: CD45 Pacific Orange™ (MHCD4530TR), CD4 PerCP-Cy®5.5 (A15858), CD20 APC, CD14 APC-Cy®7 (A15453), CD19 Pacific Green™ (C11210), CD3 Alexa Fluor® 700 (CD0329), HLA-DR PE-Cy®7 (A18558), CD8 Pacific Blue™ (MHCD0828), CD33 FITC (A16185), and CD11c PE (A18674) using the Attune® NxT Acoustic Focusing Cytometer with 405 nm excitation and 440/50 emission filter (Pacific Blue™), 512/25 emission filter (Pacific Green™), and 603/48 emission filter (Pacific Orange™); 488 nm excitation and 530/30 emission filter (FITC) and 695/50 emission filter (PerCP-Cy®5.5); 561 nm excitation and 585/16 emission filter (PE) and 780/60 emission filter (PE-Cy®7); 637 nm excitation and 660/20 emission filter (APC), 720/30 emission filter (Alexa Fluor®700), and 780/60 emission filter (APC-Cy®7). Within the CD3 negative CD45 positive gate, B cells can be identified based on expression of CD19 and CD20. Conventional dendritic cells in peripheral blood are generally negative for T and B cell lineage markers and co-express the integrin CD11c and HLA-DR. Values noted are a percent of the parent CD19 negative CD20 negative gate (top value, no parenthesis) or percent of FSC/SSC lymphocyte gate (bottom value, in parentheses).
|Tested species reactivity||Human|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Mouse / IgG1|
|Storage buffer||PBS with 4mg/ml BSA|
|Contains||0.1% sodium azide|
|Storage Conditions||4° C, store in dark|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||Assay-Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Our Pacific Orange™ dye is optimally excited by the violet laser, and it is recommended that a 575, 585 or 600 nm band pass filter be used for optimal detection. Pacific Blue™ and Pacific Orange™ dye conjugates can be simultaneously excited at 405 nm and emit at 455 nm and 551 nm, respectively, facilitating two-color analysis.
CD45 (LCA, leukocyte common antigen) is a receptor-type protein tyrosine phosphatase ubiquitously expressed in all nucleated hematopoietic cells, comprising approximately 10% of all surface proteins in lymphocytes. CD45 glycoprotein is crucial in lymphocyte development and antigen signaling, serving as an important regulator of Src-family kinases. CD45 protein exists as multiple isoforms as a result of alternative splicing; these isoforms differ in their extracellular domains, whereas they share identical transmembrane and cytoplasmic domains. These isoforms differ in their ability to translocate into the glycosphingolipid-enriched membrane domains and their expression depends on cell type and physiological state of the cell. Besides the role in immunoreceptor signaling, CD45 is important in promoting cell survival by modulating integrin-mediated signal transduction pathway and is also involved in DNA fragmentation during apoptosis. CD45RA is an isoform of the CD45 complex and has restricted expression between different subtypes of lymphoid cells.
Analyte Specific Reagent
Genetic defects in PI3K¿ affect B-cell differentiation and maturation leading to hypogammaglobulineamia and recurrent infections.
MHCD4530TR was used in flow cytometry to discuss how mutations in PIK3CD and PIK3R1 cause activated PI3K-delta syndrome
|Wentink M,Dalm V,Lankester AC,van Schouwenburg PA,Schölvinck L,Kalina T,Zachova R,Sediva A,Lambeck A,Pico-Knijnenburg I,van Dongen JJ,Pac M,Bernatowska E,van Hagen M,Driessen G,van der Burg M||Clinical immunology (Orlando, Fla.) (176:77)||2017|
Dynamic change in natural killer cell type in the human ocular mucosa in situ as means of immune evasion by adenovirus infection.
MHCD4530TR was used in flow cytometry to investigate the dynamics and characteristics of natural killer cell types in the human ocular mucosal surface in situ during infection with group D human adenoviruses.
|Yawata N,Selva KJ,Liu YC,Tan KP,Lee AW,Siak J,Lan W,Vania M,Arundhati A,Tong L,Li J,Mehta JS,Yawata M||Mucosal immunology (9:159)||2016|
Characterization of human antiviral adaptive immune responses during hepatotropic virus infection in HLA-transgenic human immune system mice.
MHCD4530TR was used in flow cytometry to investigate the role of HLA molecules during infection using humanized mice
|Billerbeck E,Horwitz JA,Labitt RN,Donovan BM,Vega K,Budell WC,Koo GC,Rice CM,Ploss A||Journal of immunology (Baltimore, Md. : 1950) (191:1753)||2013|
FcRL5 as a target of antibody-drug conjugates for the treatment of multiple myeloma.
MHCD4530TR was used in flow cytometry to investigate the use of FcRL5 antibody-drug conjugates as a treatment for multiple myeloma.
|Elkins K,Zheng B,Go M,Slaga D,Du C,Scales SJ,Yu SF,McBride J,de Tute R,Rawstron A,Jack AS,Ebens A,Polson AG||Molecular cancer therapeutics (11:2222)||2012|
Rapid ex vivo isolation and long-term culture of human Th17 cells.
MHCD4530TR was used in flow cytometry to describe a novel technique to isolate viable Th17 cells based on their ability to secrete IL-17.
|Streeck H,Cohen KW,Jolin JS,Brockman MA,Meier A,Power KA,Waring MT,Alter G,Altfeld M||Journal of immunological methods (333:115)||2008|
Analysis of CD10+ hairy cell leukemia.
MHCD4530TR was used in flow cytometry to examine CD10 expression in hairy cell leukemia.
|Jasionowski TM,Hartung L,Greenwood JH,Perkins SL,Bahler DW||American journal of clinical pathology (120:228)||2003|
Prospective isolation of human clonogenic common myeloid progenitors.
MHCD4530TR was used in flow cytometry to report how to isolate highly purified hematopoietic intermediates.
|Manz MG,Miyamoto T,Akashi K,Weissman IL||Proceedings of the National Academy of Sciences of the United States of America (99:11872)||2002|
Precise CD4 T-cell counting using red diode laser excitation: for richer, for poorer.
MHCD4530TR was used in flow cytometry to discuss the high cost of measuring CD4 T cells by flow cytometry.
|Janossy G,Jani IV,Kahan M,Barnett D,Mandy F,Shapiro H||Cytometry (50:78)||2002|
Short-term, serum-free, static culture of cord blood-derived CD34+ cells: effects of FLT3-L and MIP-1alpha on in vitro expansion of hematopoietic progenitor cells.
MHCD4530TR was used in flow cytometry to identify the optimal condition for the generation of committed progenitors from cord blood cells without affecting the stem cell compartment.
|Capmany G,Querol S,Cancelas JA,García J||Haematologica (84:675)||1999|
Illegitimate transcription: its use for studying genetic abnormalities in lymphoblastoid cells from patients with Glanzmann thrombasthenia.
MHCD4530TR was used in flow cytometry to study RNA transcripts in patients with Glanzmann thrombasthenia.
|Negrier C,Vinciguerra C,Attali O,Grenier C,Larcher ME,Dechavanne M||British journal of haematology (100:33)||1998|
Growth of human hematopoietic cells in immunodeficient mice conditioned with cyclophosphamide and busulfan.
MHCD4530TR was used in immunocytochemistry to report that human hematopoietic cells survive and proliferate in severe combined immunodeficient mice treated with busulfan and cyclophosphamide.
|Basch RS,Quito FL,Beh J,Hirst JA||Stem cells (Dayton, Ohio) (15:314)||1997|