Flow cytometry was performed using single-cell suspension, which was prepared as described with minor modifications.50 (link), 51 (link), 52 (link) Briefly, mouse hearts were minced and digested with 1.6 mg/ml collagenase IA (Sigma Aldrich, Tokyo, Japan) and 200 μg/ml DNase I (Roche, Indianapolis, IN, USA) in PBS at 37 °C for 50 min. Then cell suspensions were filtered and collected at 300 × g for 10 min. Mouse spleens were removed, triturated in PBS at 4 °C with the end of a 3-ml syringe and filtered through nylon mesh (BD Biosciences, San Jose, CA, USA). The cell suspension was centrifuged at 300 × g for 10 min at 4 °C. Red blood cells were lysed with ACK lysis buffer, and the splenocytes were washed with PBS. Flow cytometry was carried out using the following antibodies: PE anti-mouse F4/80 (Biolegend, San Diego, CA, USA), PE-cy7 anti-mouse Ly6G, PerCP-Cy5.5 anti-mouse CD45.2, PE-CF594 anti-mouse CD3e, Alexa 488 anti-mouse CD11b, APC-cy7 anti-mouse CD11b, V450 anti-mouse Ly6C, Alexa 488 anti-mouse CD31 and PE anti-mouse PDGFα (all from BD Biosciences). Flow cytometry data were acquired using BD LSRFortessa (BD Biosciences) and analyzed by BD FACSDiva software (BD Biosciences). FACS sorting was performed on a BD FACS Aria II System (BD Biosciences).
Pe cy7 anti mouse ly6g
Manufactured by BD
Sourced in United States
PE-cy7 anti-mouse Ly6G is a fluorochrome-conjugated antibody that binds to the Ly6G antigen expressed on mouse granulocytes. It can be used for the identification and enumeration of these cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Nylon mesh, by BD (1 mentions)
Pe cf594 anti mouse cd3e, by BD (1 mentions)
Apc cy7 anti mouse cd11b, by BD (1 mentions)
Pe anti mouse f4 80, by BioLegend (1 mentions)
Facsaria 2 system, by BD (1 mentions)
Collagenase ia, by Merck Group (1 mentions)
Lsrfortessa, by BD (1 mentions)
Dnase 1, by Roche (1 mentions)
Facsdiva software, by BD (1 mentions)
Apc anti mouse f4 80 clone bm8, by BD (1 mentions)
Horizon fixable viability stain 700, by BD (1 mentions)
Zombie aqua fixable viability kit, by Thermo Fisher Scientific (1 mentions)
Fitc anti mouse ter 119, by BioLegend (1 mentions)
Cytoflex benchtop flow cytometer, by Beckman Coulter (1 mentions)
Skeletal muscle dissociation kit, by Miltenyi Biotec (1 mentions)
2 protocols using pe cy7 anti mouse ly6g
1
Flow Cytometry Analysis of Immune Cell Populations
2
Multiparametric Flow Cytometry of Muscle Leukocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total leukocytes from naïve C57BL/6 muscle were isolated as described above and multiple populations identified using a CytoFLEX benchtop flow cytometer equipped with 405, 488, 561, and 637 nm solid phase lasers (Beckman Coulter Life Sciences, Brea, CA, USA) using the following fluorescent mAb (BioLegend):
FITC anti-mouse Ter119, PE/Cyanine7 anti-mouse Ly-6A/E (Sca-1), PE anti-mouse CD284 (TLR4), APC/Cy7 anti-mouse CD45, Brilliant Violet 421 anti-mouse CD31, and Zombie Aqua Fixable Viability Kit, as well as eFluor 660 anti-mouse/human CD34 (Thermo Fisher Scientific) (antibodies listed in Supplemental Table S2). At 4 days post-surgery, total leukocytes from muscle were isolated using a skeletal muscle dissociation kit (Miltenyi Biotech). Total leukocytes were subsequently discriminated into multiple populations using a CytoFLEX benchtop flow cytometer using the following mAb (BioLegend; outlined in Supplemental Table S1): FITC antimouse Ter119, FITC anti-mouse B220, FITC anti-mouse CD3ε, APC anti-mouse F4/80 (clone BM8), PE/Cy7 anti-mouse Ly6G, Brilliant Violet 510 anti-mouse/human CD11b, Brilliant Violet 785 anti-mouse CD45, and BD Horizon Fixable Viability Stain 700 (Supplemental Table S1). Files were subsequently analyzed with Flow Jo software version 10.5 (Becton Dickinson & Company, San Diego, CA, USA).
FITC anti-mouse Ter119, PE/Cyanine7 anti-mouse Ly-6A/E (Sca-1), PE anti-mouse CD284 (TLR4), APC/Cy7 anti-mouse CD45, Brilliant Violet 421 anti-mouse CD31, and Zombie Aqua Fixable Viability Kit, as well as eFluor 660 anti-mouse/human CD34 (Thermo Fisher Scientific) (antibodies listed in Supplemental Table S2). At 4 days post-surgery, total leukocytes from muscle were isolated using a skeletal muscle dissociation kit (Miltenyi Biotech). Total leukocytes were subsequently discriminated into multiple populations using a CytoFLEX benchtop flow cytometer using the following mAb (BioLegend; outlined in Supplemental Table S1): FITC antimouse Ter119, FITC anti-mouse B220, FITC anti-mouse CD3ε, APC anti-mouse F4/80 (clone BM8), PE/Cy7 anti-mouse Ly6G, Brilliant Violet 510 anti-mouse/human CD11b, Brilliant Violet 785 anti-mouse CD45, and BD Horizon Fixable Viability Stain 700 (Supplemental Table S1). Files were subsequently analyzed with Flow Jo software version 10.5 (Becton Dickinson & Company, San Diego, CA, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!