Erythroclear red blood cell depletion reagent kit

Manufactured by STEMCELL

Erythroclear Red Blood Cell Depletion Reagent Kit is a laboratory product designed to deplete red blood cells from biological samples. It serves as a tool for researchers to isolate and enrich other cell types of interest from complex mixtures.

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Lab products found in correlation

Chromium single cell 3 library and gel bead kit v3, by 10x Genomics (1 mentions) Tapestation 4200, by Agilent Technologies (1 mentions) Novaseq 6000, by Illumina (1 mentions) Dynabeads myone silane beads, by Thermo Fisher Scientific (1 mentions) 96 deep well reaction module, by Bio-Rad (1 mentions) Ficoll hypaque gradient, by GE Healthcare (1 mentions) Easysep direct human neutrophil isolation kit, by STEMCELL (1 mentions)

2 protocols using erythroclear red blood cell depletion reagent kit

Single-Cell RNA Sequencing Protocol

Cited 1 time

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The following procedures were previously described. For single-cell sequencing, white blood cells were isolated with the Erythroclear Red Blood Cell Depletion Reagent Kit (STEMCELL Technologies, catalog no. 01738), according to the manufacturer’s protocol. Cells were washed with PBS with 0.02% bovine serum albumin. To obtain single-cell gel beads in emulsion (GEMs), we resuspended cells at a concentration of 1000 cells/μl and loaded the mix on the Chromium Comptroller Instrument (10x Genomics). Single-cell cDNAs and libraries were prepared with Chromium Single-Cell 3′ Library and Gel Bead Kit v3.1 (10x Genomics, catalog no. 1000121). Briefly, GEM-RT incubation was performed in a C1000 Touch thermal cycler with 96-deep well reaction module (Bio-Rad, catalog no. 1851197). Single-strand cDNAs were purified with DynaBeads MyOne Silane Beads (Thermo Fisher Scientific, catalog no. 37002D) and amplified with the C1000 cycler. Amplified cDNA products were cleaned with 0.6X DynaBeads MyOne Silane Beads (Thermo Fisher Scientific, catalog no. 37002D). Quality and quantity of the cDNAs were assessed on 4200 TapeStation (Agilent Technologies) with High Sensitivity D5000 DNA ScreenTape (Agilent, catalog no. 5067-5592). Libraries were diluted to the same molarity and pooled for sequencing on NovaSeq6000 (Illumina) sequencers (23 (link)).

Wigerblad G., Warner S.A., Ramos-Benitez M.J., Kardava L., Tian X., Miao R., Reger R., Chakraborty M., Wong S., Kanthi Y., Suffredini A.F., Dell’Orso S., Brooks S., King C., Shlobin O., Nathan S.D., Cohen J., Moir S., Childs R.W., Kaplan M.J., Chertow D.S, & Strich J.R. (2023). Spleen tyrosine kinase inhibition restores myeloid homeostasis in COVID-19. Science Advances, 9(1), eade8272.

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Isolation of Human Neutrophils and Granulocytes

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Human venous peripheral blood samples from healthy donors were obtained from the Department of Transfusion Medicine at the National Institutes of Health Clinical Center. For neutrophil purification, whole blood samples were collected in vacutainer glass blood collection tubes with acid citrate dextrose (ACD). Neutrophils were isolated with the EasySep Direct Human Neutrophil Isolation Kit (STEMCELL Technologies; cat. no. 19666).
For granulocyte purification, whole blood was collected in heparinized tubes. Granulocytes were isolated by dextran sedimentation of RBC pellets as previously described (18 ). Briefly, cells were first layered on a Ficoll/Hypaque gradient (GE Healthcare; cat. no. 17144003). The granulocyte/RBC fraction was then enriched by dextran sedimentation followed by RBC lysis using hypotonic solution. Granulocytes were then washed with phosphate-buffered saline (PBS). For white blood cell purification, whole blood samples were collected in heparinized tubes. White blood cells were isolated with the Erythroclear Red Blood Cell Depletion Reagent Kit (STEMCELL Technologies; cat. no. 01738).

Wigerblad G., Cao Q., Brooks S., Naz F., Gadkari M., Jiang K., Gupta S., O’Neil L., Dell’Orso S., Kaplan M.J, & Franco L.M. (2022). Single-Cell Analysis Reveals the Range of Transcriptional States of Circulating Human Neutrophils. The Journal of Immunology Author Choice, 209(4), 772-782.

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