Apc conjugated cd235a

Manufactured by BD

APC-conjugated CD235a is a cell surface marker used to identify and analyze erythroid cells in flow cytometry applications. It binds to the CD235a (Glycophorin A) antigen, which is expressed on the surface of red blood cells and their precursors. The APC (Allophycocyanin) fluorochrome is conjugated to the CD235a antibody, allowing for detection and quantification of erythroid cells in samples.

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Lab products found in correlation

May gr nwald giemsa stain, by Merck Group (1 mentions) Diva software version 6, by BD (1 mentions) Facsaria, by BD (1 mentions) Sytox blue, by Thermo Fisher Scientific (1 mentions) Axioskop 40, by Zeiss (1 mentions) Alexa fluor 488 labeled donkey anti goat igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 labeled donkey anti rabbit igg, by Thermo Fisher Scientific (1 mentions)

2 protocols using apc conjugated cd235a

Cytospin and Flow Cytometry Analysis

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Cytospin preparations were obtained by cytocentrifugation (Shandon, Astmoor, UK) and stained with May-Grünwald-Giemsa stain (Sigma). Slides were observed with a Axioskop 40 microscope (Zeiss, Oberkochen, Germany) and images acquired with a CoolSNAP cf digital CCD digital camera (Photometrics Scientific, Tucson, AZ). For flow cytometry determinations of erythroid cells, samples were labeled with APC-conjugated CD235a and FITC-conjugated CD36 (Becton Dickinson Biosciences, Franklin Lakes, NJ). Dead cells were identified by staining with Sytox blue (1 μM) (Life Technologies, Waltham, MA). Cell fluorescence was analyzed with a FACSAria (Becton Dickinson Biosciences).
Cell Cycle Analyses: MNC (1.0–3.0×10⁶) cultured in HEMA for 24, 48 and 72h with or without SB431542 (26 μM) were labelled with phycoerythrin-cyanin 7 (PE-Cy7)-conjugated CD34 or appropriate isotype controls (Becton Dickinson Biosciences), fixed overnight with paraformaldehyde (8% v/v) and then permeabilized with Triton X-100 (0.25% v/v). Permeabilized cells were labelled with phycoerythrin Ki-67 (Becton Dickinson Biosciences) and DAPI and analysed with the FACS Aria (Becton Dickinson Biosciences). Results were analysed with the Diva software version 6.1.3 (Becton Dickinson Biosciences).

Ceglia I., Dueck A.C., Masiello F., Martelli F., He W., Federici G., Petricoin EF I.I.I., Zeuner A., Iancu-Rubin C., Weinberg R., Hoffman R., Mascarenhas J, & Migliaccio A.R. (2016). Pre-clinical rationale for TGF-β inhibition as a therapeutic target for the treatment of myelofibrosis. Experimental hematology, 44(12), 1138-1155.e4.

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Characterizing Erythroid Cell Expression

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CD71 and CD235a positive cells were detected with FITC-conjugated CD71 mAb (1:100, Becton, Dickinson) and APC-conjugated CD235a (glycophorin A) mAb (1:500, Beckton Dickinson). Analyses of MCR expression levels were undertaken according to a specific procedure. In brief, cells were fixed with 4% paraformaldehyde and blocked with Human TruStain FcX (Fc Receptor Blocking Solution, Bio-legend, CA, USA) for 15 min. The cells were incubated with goat anti-human MC5R pAb (25 μg/ml; Abcam) in Tris-HCl buffer with 0.25% Triton X-100 for 1 h and then incubated with secondary Abs [Alexa Fluor 488-labeled donkey anti-rabbit IgG (Molecular Probes) or Alexa Fluor 488-labeled donkey anti-goat IgG] for 30 min at RT.

Simamura E., Arikawa T., Ikeda T., Shimada H., Shoji H., Masuta H., Nakajima Y., Otani H., Yonekura H, & Hatta T. (2015). Melanocortins Contribute to Sequential Differentiation and Enucleation of Human Erythroblasts via Melanocortin Receptors 1, 2 and 5. PLoS ONE, 10(4), e0123232.

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