Primary antibodies used for IHC and flow cytometry were as follows: c-Kit (2B8; eBioscience), CD16/32 (93; eBioscience), VE-cadherin (eBioscience), c-Kit (R&D Systems), Sca-1 (E13-161.7; BioLegend), CD48 (HM48-1; BioLegend), CD150 (TC15-12F12.2; BioLegend), IL-7Rα (SB/199; BioLegend), endoglin (MJ7/18; BioLegend), CD4 (L3T4; BD), CD8 (53-6.72; BD), B220 (RA3-6B2; BD), TER-119 (BD), Gr-1 (RB6-8C5; BD), CD34 (RAM34; BD), Mac-1 (M/70; BD), Flt-3 (A2F10.1; BD), CD41 (MWReg30; BD), CD45.2 (104; BD), CD45.1 (A20; BD), GPIbα (Xia.G5; Emfret), Clec2 (AbD Serotec), and Thpo (Bioss). Secondary antibodies for IHC were Alexa Fluor 488–conjugated IgGs (Molecular Probes) or Cy3/Cy5/DyLight549/DyLight649-conjugated IgGs (Jackson ImmunoResearch Laboratories, Inc.). IHC specimens were treated with DAPI (Molecular Probes) for nuclear staining. Stimulatory rabbit anti–mouse CLEC-2 antibody was a gift of K. Suzuki-Inoue.
Cd34 ram34
Manufactured by BD
Sourced in Finland
The CD34 (RAM34) is a lab equipment product used for the detection and analysis of CD34 antigen, a cell surface marker expressed on hematopoietic stem and progenitor cells. The product is designed to facilitate the identification and enumeration of CD34+ cells in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Gr 1 rb6 8c5, by BD (2 mentions)
Cd48 hm48 1, by BioLegend (2 mentions)
Cd150 tc15 12f12.2, by BioLegend (2 mentions)
Flt 3 a2f10.1, by BD (2 mentions)
Ter119, by BD (2 mentions)
B220 ra3 6b2, by BD (2 mentions)
Cd135 a2f10, by Thermo Fisher Scientific (2 mentions)
C kit 2b8, by Thermo Fisher Scientific (1 mentions)
Cd16 32, by Thermo Fisher Scientific (1 mentions)
Ve cadherin, by Thermo Fisher Scientific (1 mentions)
Sca 1 e13 161.7, by BioLegend (1 mentions)
Cd4 l3t4, by BD (1 mentions)
Cd8 53 6.72, by BD (1 mentions)
Mac 1 m 70, by BD (1 mentions)
Cd41 mwreg30, by BD (1 mentions)
Cd45.1 a20, by BD (1 mentions)
Cy3 cy5 dylight549 dylight649 conjugated iggs, by Jackson ImmunoResearch (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
C kit, by R&D Systems (1 mentions)
Tcrβ h57 597, by BD (1 mentions)
5 protocols using cd34 ram34
1
Immunophenotyping of Hematopoietic Cells
2
Multiparameter Flow Cytometry of Mouse Hematopoietic Cells
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The following anti-mouse mAbs were used: Sca-1 (E13161.7, produced in house), c-Kit (ACK2, produced in house), Flt3 (A2F10.1; BD Pharmingen), IL-7Rα (B12-1; eBioscience, Bioof), Ly6C (5075-3.6), Ly6D (49-H4, BD Pharmingen), CD19 (ID3; BD Pharmingen, eBioscience), B220 (RA3-6B2; BD Pharmingen, eBioscience), IgM (331.12, BD Pharmingen), NK1.1 (PK136, BD Pharmingen), CD49b (HMα2, BD Pharmingen), TCRβ (H57-597, BD Pharmingen), CD45.1 (A20, eBioscience), and CD34 (RAM34; BD Pharmingen). Anti-rat immunoglobulin-phycoerythrin and PECy7-streptavidin (BD Pharmingen) were used as secondary detection reagents.
Single cell suspensions were prepared in balanced salt solution with 2% (v/v) fetal calf serum. Cell staining was on ice for 30 min with fluorescent or biotin conjugated antibodies and the samples were processed on an LSRII or LSRFortessa flow cytometer (BD Biosciences). Propidium iodide exclusion was used to determine cell viability. Data were analyzed using FlowJo software (Treestar Inc.).
Single cell suspensions were prepared in balanced salt solution with 2% (v/v) fetal calf serum. Cell staining was on ice for 30 min with fluorescent or biotin conjugated antibodies and the samples were processed on an LSRII or LSRFortessa flow cytometer (BD Biosciences). Propidium iodide exclusion was used to determine cell viability. Data were analyzed using FlowJo software (Treestar Inc.).
3
Multiparametric Flow Cytometry of Murine Spleen Cells
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For flow cytometric analyses, spleen cells from mice were harvested on the 7th day post-infection, suspended in 50 μl of FACS buffer (PBS, 2% FCS) and surface stained at 4 °C with anti-mouse CD4 (GK1.5, Cat#17-0041-82) -CD8 (53-6.7 Cat#11-0081-82), -CD19 (1D3, Cat#17 0193-82), -CD29 (HM-Bta1-1 Cat#12-0291-82), -CD3 (2C11 Cat#17-0032-82), -Sca-1 (Ly-6A/E Cta#11-5981-82) PD-1 (MIH4 Cat#12-9969-42) were purchased from e-Biosciences and -CD34 (RAM34 Cat#551387 BD Biosciences) Stained cells were acquired with a FACS Fortessa (BD Biosciences) and analysed by Flow Jo (Tree star) or cyflogic (CyFlo Ltd, Finland) software.
4
Multicolor Flow Cytometry for Bone Marrow and Spleen Progenitors
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For assessment of bone marrow and spleen progenitor cells by flow cytometry, single-cell suspensions of freshly prepared bone marrow and spleen were incubated with the following anti-mouse monoclonal antibodies: Ter119 (Ly-76, BD Biosciences), CD71 (C2, BD Biosciences), CD34 (RAM34, BD Biosciences), CD135 (A2F10, eBioscience), CD127 (A7R34, eBioscience), c-Kit (ACK2, eBioscience), SCA-1 (LY-6A/E, BD Biosciences), eFluor520 (eBioscience), and 7-AAD (eBioscience). Lineage (lin) staining included anti-mouse Ter119 (BD Biosciences), GR-1 (RB6-8C5, BD Biosciences), B220 (RA3-6B2, BD Biosciences), CD4 (RM4-5, BD Biosciences), CD8 (53–6.7, BD Biosciences), and CD11b (M1/70, BD Biosciences) antibodies [22 (link)–24 (link)]. Multicolor data acquisition for bone marrow progenitor subsets was performed on a FACSCanto II flow cytometer, and data were analyzed with FlowJo 7.6.1.
5
Multiparameter Phenotyping of Hematopoietic Cells
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Single-cell suspensions from spleen, blood, BM, or BALF were washed in PBS containing 5 mM ethylenediaminetetraacetic acid and 0.5% BSA. Cells were incubated for 15 min with Fc receptor block (clone 2.4G2; BD) before antibody staining. The following antibodies were used for FACS staining and sorting: biotinylated antibodies for lineage-negative staining-biotin lineage staining cocktail includes CD3, B220, GR-1, CD11b, and Ter119, as well as NK1.1, CD11c, and DX-5 (BD), were added to the staining cocktail; CD45.2 (104), cKit (2B8), and CD34 (Ram34) were purchased from BD; CD16/32 (93), Sca-1 (D7), and CD135 (A2F10) were purchased from eBioscience; CD45.1 (A20), CD48 (HM48-1), and CD150 (TC15-12F12.2) were purchased from BioLegend. Aqua Fluorescent reactive dye (Life Technologies) was used to discriminate viable from dead cells. Intracellular BrdU and 7AAD for cell cycle analysis using the FITC BrdU Flow kit (BD) was performed per the manufacturer’s protocol. Flow cytometry data was analyzed using FlowJo software (Tree Star).
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