Human embryonic kidney cells (HEK293), Mac-1-expressing HEK293 cells (Mac-1-HEK293), LFA-1 (CD11a/CD18) and HEK293 cells expressing the αM subunit lacking the αMI-domain [20 (link)] were maintained in DMEM (Mediatech Inc, Manassas, VA) supplemented with 10 % fetal bovine serum and antibiotics. U937 monocytic cells were grown in RPMI containing 10 % fetal bovine serum and antibiotics. Neutrophils were isolated under sterile conditions from human peripheral blood obtained from consenting volunteers as described [59 (link)]. Peritoneal macrophages were isolated from wild-type and Mac-1-deficient mice (The Jackson Laboratories) 3 days after the injection of 4 % thioglycollate (TG). Macrophages were separated from lymphocytes using a mouse PE selection kit (STEMCELL Technologies, Vancouver, BC, Canada).
Mouse pe selection kit
Manufactured by STEMCELL
Sourced in Canada
The Mouse PE Selection Kit is a laboratory product designed to perform positive selection of mouse cells expressing the PE (Phycoerythrin) marker. It provides a simple and effective way to isolate and enrich cell populations of interest based on their PE expression.
Automatically generated - may contain errors
Lab products found in correlation
Anti sca 1 pe, by BD (1 mentions)
Superscript 3 first strand, by Thermo Fisher Scientific (1 mentions)
Power sybr green master mix, by Thermo Fisher Scientific (1 mentions)
Mirneasy mini kit, by Qiagen (1 mentions)
Pe anti mouse gr 1, by BioLegend (1 mentions)
Macs cell separation system, by Miltenyi Biotec (1 mentions)
3 protocols using mouse pe selection kit
1
Cell Isolation and Culture Protocols
2
Quantification of Gene Expression in Fancd2 Knockout
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KSL bone marrow or ASL FL cells were sorted by FACS on BD InFlux. SCA1+ cells were isolated from bone marrow and FL using the Mouse PE Selection Kit (STEMCELL Technologies) with PE anti-SCA1 (BD 553336), and placed in Qiazol. Total RNA was purified with the miRNeasy Mini Kit (Qiagen). mRNA was converted to cDNA using SuperScript III First Strand (Invitrogen). Real-time qPCR was performed on StepOne Plus using Power SYBR Green Master Mix (Applied Biosystems). Primer sequences are described in Table S1 . Fancd2−/− and WT samples were normalized to β-actin, and Fancd2−/− gene expression was compared with WT.
3
Gr1+ cell migration assay
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Cell separation was conducted using an MACS cell-separation system (Miltenyi Biotec) using PE anti-mouse Gr1 (catalogue #108407; Biolegend) and a mouse PE selection kit (catalogue #18554; STEMCELL Technologies).
In Vitro Chemotaxis Assay Cell migration was detected using a transwell system. Briefly, cells were incubated for 3 hr at 37 C with specified chemokines or a soluble receptor (CCR5-Ig), and cell counts were determined by FACS. The migration index represents the ratio of cells with supplements compared to the non-treated control well.
In Vitro Chemotaxis Assay Cell migration was detected using a transwell system. Briefly, cells were incubated for 3 hr at 37 C with specified chemokines or a soluble receptor (CCR5-Ig), and cell counts were determined by FACS. The migration index represents the ratio of cells with supplements compared to the non-treated control well.
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