CD11b (M1/70), and Gr-1 (RB6-8C5) antibodies from eBioscience were used for flow cytometry. Flow cytometry data were acquired on an upgraded eight-color FACScan and analyzed by using FlowJo software (Tree Star).
Gr 1 rb6 8c5
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Gr-1 (RB6-8C5) is a monoclonal antibody that targets the Gr-1 antigen, also known as Ly-6G and Ly-6C. It is commonly used in flow cytometry and cell sorting applications to identify and isolate granulocytic cell populations, such as neutrophils and myeloid-derived suppressor cells (MDSCs). The antibody recognizes a cell surface marker that is expressed on these cell types.
Automatically generated - may contain errors
Lab products found in correlation
Cd11b m1 70, by Thermo Fisher Scientific (5 mentions)
F4 80 bm8, by Thermo Fisher Scientific (3 mentions)
B220 ra3 6b2, by BD (3 mentions)
Facscalibur, by BD (3 mentions)
Cd117 2b8, by BD (2 mentions)
Fcεriα mar 1, by Thermo Fisher Scientific (2 mentions)
Cd49b dx5, by BioLegend (2 mentions)
Ccr3 tg14, by BioLegend (2 mentions)
Cd3ε 145 2c11, by Thermo Fisher Scientific (2 mentions)
Cd16 32 2.4g2, by BioXCell (2 mentions)
Propidium iodide, by BD (2 mentions)
Lsrii flow cytometer, by BD (2 mentions)
Ter 119 ter119, by Thermo Fisher Scientific (2 mentions)
Cd8 53 6, by Thermo Fisher Scientific (2 mentions)
B220 ra3 6b2, by Thermo Fisher Scientific (2 mentions)
Mac 1 m1 70, by Thermo Fisher Scientific (2 mentions)
Ter119, by Thermo Fisher Scientific (2 mentions)
Cd11b m1 70, by BioLegend (2 mentions)
Siglecf e50 2440, by BD (2 mentions)
Lsr 2 flow cytometer, by BD (2 mentions)
23 protocols using gr 1 rb6 8c5
1
Flow cytometry analysis of M1/70 and RB6-8C5
2
Comprehensive Immune Cell Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Red blood cell indices were measured by an automated hematology analyzer. Leukocyte populations in blood were assessed by flow cytometry as follows: neutrophils Gr-1+, T cells CD3+, B cells B220+, monocytes F4/80+ Gr-1−, eosinophils CCR3+ SSChi, basophils FcεRIα+ CD49b+. Peritoneal cells were isolated by peritoneal lavage with PBS. MCs were identified by staining peritoneal cells for FcεRIα and CD117 (c-kit). RBCs were lysed in ACK buffer (0.15 M NH4Cl, 1 mM KHCO3, 0.1mM EDTA). Cells were blocked with anti-CD16/32 for 10 min on ice, before staining with antibodies for flow cytometry for 30 min on ice. Propidium iodide (BD Biosciences, San Jose, California, USA) was used to exclude dead cells from analysis. Antibodies used were: FcεRIα (MAR-1; eBiosciences, San Diego, California, USA), CD117 (2B8; BD Biosciences), CD49b (DX5; Biolegend, San Diego, California, USA), CCR3 (TG14; Biolegend), B220 (RA3–6B2, BD Biosciences), CD3ε (145–2C11; eBiosciences), Gr-1 (RB6–8C5; eBiosciences), F4/80 (BM8; eBiosciences), CD16/32 (2.4G2, Bio X Cell, West Lebanon, New Hampshire, USA).
3
Flow Cytometry Immunophenotyping of Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used for cell surface staining: CD11b (M1/70), CD19 (6D5), CD3 (17A2), and Gr-1 (RB6-8C5; all from eBiosciences). Cells were stained, run on LSRII flow cytometer (BD Biosciences), and analyzed with FlowJo software version 10.2 (Flowjo LLC). After removal of doublets, B cells were gated as CD19+ and T cells as CD3+; CD11b+ and PMN (CD11b+Gr1+) were gated on cells negative for both CD3 and CD19.
4
Isolation of HSC (LK+S+ cells) from Bone Marrow
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For HSC (LK+S+ cells) isolation, BM cells were firstly enriched for c-Kit expression by immuno-selection with CD117 conjugated micro-magnetic beads (Miltenyi Biotec) according to the manufacturer’s instructions. Enriched cells were then stained with PE-cy7 conjugated with a mixture of lineage antibodies (anti-CD3 145-2C11, Mac-1 M1/70, Gr-1 RB6-8C5, CD4 GK1.5, B220 RA3-6B2, CD8 53-6.7, Ter-119 TER119; all were purchased from e-Bioscience), PE conjugated Sca-1 (D7, e-Bioscience), APC conjugated c-Kit (2B8, e-Bioscience) and Percp-cy5.5 conjugated CD45.2 (104, e-Bioscience). Normal hematopoietic and leukemic cells were sorted by CD45.2 and GFP expression, respectively; and 4′,6-diamidino-2-phenylindole (DAPI) was used to exclude dead cells during the sorting procedure.
5
Isolation of Murine Hematopoietic Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Single-cell suspensions of whole BM cells were labeled by incubation on ice for 15 min using a cocktail of the following antibodies: PE-conjugated anti-mouse CD3ε (145-2C11; BD Biosciences), CD4 (L3T4; BD Biosciences), CD8α (53-6.7; BD Biosciences), B220 (RA3-6B2; BD Biosciences), Mac-1 (M1/70; BD Biosciences), Gr-1 (RB6-8C5; eBioscience), and Ter119 (TER-119, eBioscience); FITC-conjugated anti-mouse Sca-1 (E13-161.7; BD Biosciences); and APC-conjugated anti-mouse c-Kit (2B8; BD Biosciences). After cells were washed with PBS, they were incubated with anti-PE microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany) at 4°C for 15 min. Lin+ cells (CD4+, CD8α+, B220+, Mac-1+, Gr-1+, Ter119+ cells) were removed by immunomagnetic selection using a MACS system (Miltenyi Biotec). These negatively selected cells were subjected to FACS using a Moflo XDP cell sorter (Beckman-Coulter) to prepare LSK cells.
6
Isolation and Analysis of Murine Lung Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were euthanized by intraperitoneal administration of pentobarbital at the indicated time points. Lungs were isolated in HBSS buffer (Sigma, St. Louis, MO), digested with type IV collagenase (Worthington Biochemical, Lakewood, NJ) for 30 min with agitation, and then processed into single-cell suspensions by forcing the digested lungs through a 70-µm nylon cell strainer (Falcon, Corning, NY). Single-cell suspensions were stained in PBS supplemented with 1% bovine serum albumin and 2 mM EDTA. Antibodies against the following immune markers were used for discrimination of immune cell types: CD45 (30-F11; BD), CD11b (M1/70; BioLegend), CD11c (HL3; BD), Ly6c (HK1.4; eBioscience), GR1 (RB6-8C5; eBioscience), major histocompatibility complex class II (MHCII) (M5/114.15.2; BD), and SiglecF (E50-2440; BD). Live cells were discriminated from dead cells by using a viability dye eFluor 520 (eBioscience). All flow cytometry data were acquired on an LSR-II flow cytometer (BD) and analyzed using FlowJo software (FlowJo LCC, Ashland, OR). Cell sorting was performed on a FACS Aria (BD).
7
Multiparametric Flow Cytometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following flow cytometric antibodies were purchased from BioLegend (BioLegend, Cambridge, UK): CD4 (GK1.5), CD45R/B220 (RA3-6B2), CD14 (Sa14-2), CD138 (281-2), CD254(IK22/5), and CD11b(M1/70). Gr-1(RB6-8C5) was purchased from eBioscience Inc. (San Diego, CA). For flow cytometry, cells were isolated from bone marrow of CIA and control mice. About 1 × 106 bone marrow cells were incubated with indicated antibody mixtures. In all staining, dead cells were excluded using an Aqua Live/Dead fixable staining reagent (Invitrogen), and doublets were excluded by FSC-A vs. FSC-H gating. Data were acquired using FACSCalibur flow cytometer (BD Biosciences) and analyzed using the Flowjo 7.6 software.
8
Comprehensive Immune Cell Profiling
9
Colorectal Cancer Induction and Immune Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Azoxymethane (AOM), retinoic acid (RA), 3-methylcholanthrene (MCA) and Giemsa stain were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dextran sodium sulfate (DSS) was purchased from MP Biomedicals (Santa Ana, CA, USA). Anti-mouse CD3ε (145-2C11) and CD4 (RM4-5) were purchased from BD Biosciences (San Jose, CA, USA). Anti-mouse CD16/CD32 (93), and Gr-1 (RB6-8C5) mAbs were purchased from eBioscience (San Diego, CA, USA). Anti-CD8α (53–6.7), Foxp3 (3G3), and 7-amino-actinomycin D (7AAD) were from Tonbo Biosciences. The components of the AIN-93M standard diet were purchased from Oriental Yeast Co., Ltd. (Tokyo, Japan).
10
Isolation and Analysis of Intestinal Leukocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The isolated ileal muscle layer was starved for 30 min in Ca2+-free Hank’s solution and then digested for 1 h with continuous stirring at 37 °C in the presence of collagenase (Worthington Type II, Wako), bovine serum albumin (BSA; Sigma-Aldrich Japan), trypsin inhibitor (Sigma-Aldrich Japan), and adenosine triphosphate (ATP; Sigma-Aldrich Japan) in Ca2+-free Hank’s solution. Leukocytes were stained with monoclonal antibodies including anti-CD45 (30-F11; 25–0451; eBioecience, San Diego, CA), 7-Aminoactinomysin (7-AAD; 559925; BD Pharmingen Japan, Tokyo, Japan), CD11b (M1/70; 11–0112; eBioscience), Gr-1 (RB6–8C5; 12–5931; eBioscience), and Ly6C (AL-21; 560596; BD Pharmingen Japan). Samples were acquired and evaluated using a FACSVerse (BD Biosciences, Tokyo, Japan). For all samples, approximately 30,000 live 7-AAD-negative cells were analysed for plot generation.
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!