Rat igg2a pe

Manufactured by Thermo Fisher Scientific

Sourced in United States

Rat IgG2a PE is a laboratory reagent used for flow cytometry applications. It is a fluorescently labeled antibody that binds to the IgG2a subclass of rat immunoglobulins. This product can be used to detect and quantify the presence of IgG2a-expressing cells in biological samples.

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Close spelling variants of this product

IgG2a (72 citations) Rat IgG2a (30 citations) Rat IgG2a Isotype Control PE (6 citations) Anti-rat IgG2a-PE (5 citations) IgG2a-PE (4 citations) Rat IgG2a K Isotype Control PE-cy7 (4 citations) Rat IgG2a K Isotype Control PE (3 citations) PE-conjugated rat IgG2a (2 citations) Phycoerythrin-conjugated (PE) rat IgG2a (2 citations) Rat IgG2a kappa-PE (1 citations)

9 protocols using rat igg2a pe

Multi-Color Flow Cytometry Immunophenotyping

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Antibody combination panels for four or five color immune-staining were as follows:

CD3PerCP, CD4 FITC, CD25APC, and rat Foxp3PE (all from eBioscience, San Diego, CA).

CD4 PercpCy5.5, HLA-DR APC, CD127 Pac Blue and rat Foxp3 PE (eBioscience, San Diego, CA).

CD4 PercpCy5.5, CCR5 FITC, CXCR4 APC (Biolegend, San Diego, CA) and Foxp3 PE.

CD4 PercpCy5.5, HIV-1 p24 FITC (Beckman Coulter, Brea, CA), Bcl-2 PE (BD Biosciences, San Jose, CA) and Foxp3 e-fluor660 (eBioscience).

CD4 PercpCy5.5, CD127 Pac Blue, Foxp3 PE, HIV-1 p24 FITC and IFN-γ Alexa Fluor 647 (Biolegend).

Isotype antibodies used included: IgG2a APC, IgG1 Pac Blue, rat IgG2a PE, IgG2a FITC, rat IgG2a AP, IgG1 FITC, IgG1 PE, IgG2a e-fluor660, and IgG1 Alexa Fluor 647.
The protocol and buffer set from eBioscience were used for all experiments involving intracellular staining. All samples were fixed and acquired within 1h of completion of staining, and analysis was by Miltenyi MACSQuant Flow cytometer.
Data were analyzed by FlowJo software (Treestar Inc, Ashland, OR) at the completion of study.

Hirsch C.S., Baseke J., Kafuluma J.L., Nserko M., Mayanja-Kizza H, & Toossi Z. (2016). Expansion and productive HIV-1 infection of Foxp3 positive CD4 T cells at pleural sites of HIV/TB co-infection. Journal of clinical & experimental immunology, 1(1), http://www.opastonline.com/wp-content/uploads/2016/11/expansion-and-productive-hiv-1-infection-of-foxp3-positive-cd4-t-cells-at-pleural-sites-of-hivtb-co-infection-jcei-16-007.pdf.

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Myeloid Cell Subset Isolation from Tumor-Bearing Livers

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Single-cell suspensions were prepared from tumor-bearing livers as previously described.^{10 (link)} Cell suspensions were stained in the presence of Mouse BD Fc Block (BD Biosciences, Oxford, UK). Antibodies used in FACS included rat anti-mouse CD11b PE-Cy7 (eBioscience, Hatfield, UK, 25-0112-82), rat anti-mouse Gr1 PE (eBioscience, 12-5931-82) and isotype controls rat IgG2a PE and rat IgG2b PE-Cy7 (eBioscience). Flow cytometry was performed using a FACSCalibur flow cytometer (BD Biosciences) and analyzed using FlowJo software version 7.2.5 (Tree Star, Ashland, OR, USA). Myeloid cells were sorted into granulocyte (CD11b⁺/Gr1^high) or monocyte/macrophage (CD11b⁺/Gr1^mid/low) subsets using a MoFlo XDP High-Speed Cell Sorter (Beckman Coulter, High Wycombe, UK) by the antibodies described above.

Lim S.Y., Yuzhalin A.E., Gordon-Weeks A.N, & Muschel R.J. (2016). Tumor-infiltrating monocytes/macrophages promote tumor invasion and migration by upregulating S100A8 and S100A9 expression in cancer cells. Oncogene, 35(44), 5735-5745.

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Tricolor Flow Cytometry of Canine Regulatory T Cells

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Tricolor flow cytometry was used to identify the influence of MPA on the population of ConA and PHA-stimulated regulatory T (Treg) cells and the expression of CD25 –a late activation marker. Surface antigens were stained using anti-canine mAbs (Table 1): rat anti-CD4:FITC (clone YKIX302.9, MCA1038F, AbD Serotec), mouse anti-CD25:eFluor 660 (clone P4A10, 50–0250, eBioscience, San Diego, USA) according to protocol of staining extracellular antigens described above [48 (link), 49 (link)]. Next, forkhead box P3 (FoxP3) expression was evaluated using a cross-reactive anti-FoxP3:PE antibody as described previously by Biller et al. using a FoxP3/Transcription Factor Staining Buffer Set (00–5523, eBioscience) [48 (link), 50 (link)]. Briefly, the samples were incubated overnight in the 1X Fixation/Permeabilization solution at 4°C in the dark. The samples were then washed in a 1X Permeabilization Buffer and incubated with the rat anti-FoxP3:PE mAb (clone FJK-16s, 12–5773, eBioscience) cross-reacting with dog cells at room temp. for 30 min (Table 1). After staining, the samples were washed again, suspended in the cytometric buffer and analyzed. Samples incubated with corresponding isotype were used as a negative control: rat IgG2a:FITC (MCA1212F, AbD Serotec), mouse IgG1:eFluor 660 (50–4714, eBioscience) and rat IgG2a:PE (12–4321, eBioscience).

Guzera M., Szulc-Dąbrowska L., Cywińska A., Archer J, & Winnicka A. (2016). In Vitro Influence of Mycophenolic Acid on Selected Parameters of Stimulated Peripheral Canine Lymphocytes. PLoS ONE, 11(5), e0154429.

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Myeloid Cell Isolation from Tumor Bearing Livers

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Single cell suspensions were prepared from tumor bearing livers as previously described (10 ). Cell suspensions were stained in the presence of Mouse BD Fc Block (BD Bioscience). Antibodies used in FACS included rat anti-mouse CD11b PE-Cy7 (eBioscience, 25-0112-82), rat anti-mouse Gr1 PE (eBioscience, 12-5931-82) and isotype controls rat IgG2a PE and rat IgG2b PE-Cy7 (eBioscience). Flow cytometry was performed using a FACSCalibur flow cytometer (BD Biosciences) and analyzed with FlowJo software version 7.2.5 (Tree Star, Ashland, OR). Myeloid cells were sorted using a MoFlo XDP High-Speed Cell Sorter (Beckman Coulter, UK) into granulocyte (CD11b⁺/Gr1^high) or monocyte/macrophage (CD11b⁺/Gr1^mid/low) subsets using the antibodies described above.

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Flow Cytometry Analysis of Mouse Immune Cells

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Single-cell suspensions of splenocytes and lymph node cells were treated with RBC lysis buffer (Sigma), and washed with FACS buffer (PBS plus 1% FBS). These cells were incubated with specific surface-binding antibodies for 30 min at 4°C. Antibodies included anti-mouse CD3e-percp-cy5.5, anti-mouse CD19-FITC, anti-mouse CD44-FITC, anti-mouse CD62L-PE, anti-mouse B220-APC, anti-mouse CD80-APC, anti-mouse CD86-APC and rat IgG₁-APC isotrol control and rat IgG_2a-PE isotrol control (all from eBioscience, San Diego, CA), anti-mouse CD138-PE (Miltenyi Biotech, Bergisch Gladbach, Germany), anti-mouse CD4-APC, anti-mouse CD8-APC (BD pharmingen). In some experiments, splenocytes were stimulated with LPS (1 μg/ml) or PBS for 24 h before stained with fluorochome-conjugated CD80, CD86 and CD19 antibodies. For JC-1 staining, splenocytes were stimulated with anti-CD3 and anti-CD28 antibodies (1 μg/ml for each, BD pharmingen) for 24 h and then incubated with 5 μg/ml JC-1 (Biotium, Hayward, CA) for 30 min at 37°C. Samples were analyzed by flow cytometry on a FACScan flow cytometer (Becton Dickinson).

Guo L., Liu W., Lu T., Guo W., Gao J., Luo Q., Wu X., Sun Y., Wu X., Shen Y, & Xu Q. (2015). Decrease of Functional Activated T and B Cells and Treatment of Glomerulonephitis in Lupus-Prone Mice Using a Natural Flavonoid Astilbin. PLoS ONE, 10(4), e0124002.

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Multiparametric analysis of murine cells

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A formulation of XH-201 and XH-202 was used in these studies. The following antibodies were purchased from eBioscience (San Diego, CA, USA): anti-mouse Ly-6A/EA (Sca-1)-PE, rat IgG2a- PE (Isotype Ctrl), CD117 (c-kit)-APC, rat IgG2b-APC (Isotype Ctrl), biotin-conjugated CD5, CD4, CD8, CD45R/B220, Ly6G/Gr-1, CD11b, Ter-119 and PerCP-conjugated streptavidin antibodies [20 (link)]. RPMI 1640 medium was purchased from Gibco (Grand Island, NY, USA). Melatonin was purchased from Tixiai Chemical Industry Co., Ltd (Shanghai, China). BD Cytofifix/Cytoperm buffer was purchased from BD Biosciences (San Diego, CA, USA). MitoSOX red mitochondrial superoxide indicator was obtained from Life Technologies (Grand Island, NY, USA). Rabbit anti-γH2AX antibody was obtained from Cell Signaling Technology (Danvers, MA, USA) and FITC-conjugated goat anti-rabbit antibodies from Abcam (Cambridge, MA, USA).

Cao J., Li H., Yuan R., Dong Y., Wu J., Wang M., Li D., Tian H, & Dong H. (2020). Protective effects of new aryl sulfone derivatives against radiation-induced hematopoietic injury. Journal of Radiation Research, 61(3), 388-398.

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Recombinant Human Growth Hormone Protocol

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Recombinant human GH was purchased from Dong-A Pharmaceutical Co., Ltd (Dalseong-Gun, Daegu, South Korea). Laminin 111 protein, primary anti-laminin antibody and secondary goat anti-rabbit-FITC conjugated antibody were purchased from Sigma-Aldrich (St. Louis, MO, USA). Monoclonal antibodies: anti-CD4 allophycocyanin (APC), anti-CD8 peridinin chlorophyll protein (PerCP), anti-CD49f phycoerythrin (PE) and rat IgG2a PE were purchased from eBioscience (San Diego, CA, USA). Fetal bovine serum (FBS), RPMI-1640 medium, l-glutamine, gentamicin, Trypsin–EDTA, phosphate buffered saline (PBS) and bovine serum albumin (BSA) were obtained from Sigma-Aldrich.

Lins M.P., de Araújo Vieira L.F., Rosa A.A, & Smaniotto S. (2016). Growth hormone in the presence of laminin modulates interaction of human thymic epithelial cells and thymocytes in vitro. Biological Research, 49(1), 37.

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Myeloid Cell Phenotyping in Skin

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Skin cell suspension from baicalin and vehicle treated mice were prepared and CD11b⁺Gr1⁺ myeloid cells were analyzed using APC labeled CD11b and PE labeled Gr1 antibodies (BD Biosciences). The percentage of cells that expressing high levels of both CD11b and Gr1 were analyzed in a flow cytometer (BD Flow cytometer (San Diego, CA), and the data were analyzed using FlowJo software v10.5.0. Rat IgG2B-APC and rat IgG2A-PE antibodies were used as isotype controls (Thermofisher Scientific Waltham, MA). The skin cell suspensions were also stained with APC labeled CD11b, FITC labeled MHCII and PE labeled TLR4 and analyzed using Flow cytometry.

Sherwani M.A., Yang K., Jani A., Abed R.A., Taufique A.K., Dosunmu T.G, & Yusuf N. (2018). Protective Effect of Baicalin Against TLR4-mediated UVA-induced Skin Inflammation. Photochemistry and photobiology, 95(2), 605-611.

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Flow Cytometric Analysis of Myeloid Cell Markers

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Expression of F4/80, Ly-6C/6G, CD11b and CD71 was determined by flow cytometry. Cells (0.5 × 10⁶) were washed with PBS, suspended in FACS buffer and blocked during 15 min at 4 °C with 10% of normal mouse serum (ThermoFisher Scientific, Erembodegem, Belgium). Then, the cells were stained with the following fluorochrome-conjugated rat anti-mouse antibodies: anti-F4/80-APC (ThermoFisher Scientific, clone BM8), anti-Ly6C/6G-eFluor^®450 (ThermoFisher Scientific, clone RB6-8C5), anti-CD11b-FITC (ThermoFisher Scientific, clone M1/70) and anti-CD71-PE (ThermoFisher Scientific, clone R17 217.1.4). Cell samples were kept unstained or incubated with the following isotype control: rat IgG2b-APC (ThermoFisher Scientific, clone eBr2A), rat IgG2b-FITC (ThermoFisher Scientific, clone eB149/10H5), rat IgG2a-PE (ThermoFisher Scientific, clone eBr2a), and rat IgG2b-eFluor^®450 (ThermoFisher Scientific, clone eB149/10H5). Cells were incubated with antibody cocktails for 30 min at 4 °C in the presence of 7-AAD. Surface marker expression was determined on a BD FACS Canto II™ flow cytometer (BD Biosciences).

Zhou Y., Hann J., Schenten V., Plançon S., Bueb J.L., Tolle F, & Bréchard S. (2021). Role of S100A8/A9 for Cytokine Secretion, Revealed in Neutrophils Derived from ER-Hoxb8 Progenitors. International Journal of Molecular Sciences, 22(16), 8845.

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