Anti cd8 ecd

Manufactured by Beckman Coulter

Sourced in United States

The Anti-CD8-ECD is a fluorescently labeled antibody that binds to the CD8 antigen expressed on the surface of certain T cells. It is designed for use in flow cytometry applications to identify and quantify CD8+ T cells.

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3 protocols using anti cd8 ecd

Comprehensive Immune Cell Profiling

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CBC and T-cell analyses were performed by Karolinska University Laboratory, Stockholm, Sweden, using Sysmex XN-9000 for CBC processing. T-cell analysis for CD4+ and CD8+ expression was performed using an Aquios CL (Beckman coulter) which utilizes a direct volumetric single‐platform method with incorporated sample preparation with a monoclonal antibody mixture (anti‐CD45‐FITC [clone B3821F4A], anti‐CD4‐RDI [clone SFCI12T4D11], anti CD8‐ECD [SFCI21thyD3], anti‐CD3‐PC5 [clone UCHT1]) Beckman Coulter.
For PBMC isolation, whole blood was sampled using BD Vacutainer^® CPT™ Mononuclear Cell Preparation Tubes and processed within 3 hours of collection. PBMCs were then isolated according to the standard procedure (centrifuged at 1500 g for 20 min at room temperature) and washed with cold PBS (440 g for 10 min at 4°C). Single cell suspensions were plated in 96-well V-bottomed plates and stained for 20 min at 4°C. The cells were incubated with Alexa Fluor647 anti-human CX3CR1 (clone: 2A9-1, BioLegend), PerCP/Cy5.5 anti-human CD192 (CCR2) (clone: K036C2, BioLegend), APC/Cy7 anti-human CD68 (clone: Y1/82A, BioLegend), PE/Cy7 anti-human CD11b (clone: ICRF44, BioLegend), Alexa Fluor488 anti-human CD16 (clone: 3G8, BioLegend) and PE anti-human CD14 (clone: 63D3, BioLegend). Cells were acquired using a Gallios flow cytometer (Beckman Coulter) and analyzed using Kaluza software (Beckman Coulter).

Hildenborg M., Kåhlin J., Granath F., Schening A., Granström A., Ebberyd A., Klevenvall L., Zetterberg H., Han J., Schlegel T.T., Harris R., Harris H.E, & Eriksson L.I. (2022). The Neuroimmune Response to Surgery – An Exploratory Study of Trauma-Induced Changes in Innate Immunity and Heart Rate Variability. Frontiers in Immunology, 13, 911744.

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Lymphocyte Subset Analysis in COVID-19 Patients

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Based on hsTnI levels, we divided COVID-19 patients into hsTnI-L and hsTnI-H groups, and healthy physical examiners were selected as the control group. Lymphocyte subsets were compared among the three groups.
Peripheral blood samples were collected in ethylenediaminetetraacetic acid K2 (EDTA-K2) collection tubes for lymphocyte subset and hemoglobin A1c detection. Antibodies, including anti-CD3-PC5, anti-CD4-RD1, anti-CD8-ECD, anti-CD45-FITC, anti-CD56-RD1, and anti-CD19-ECD, were purchased from Beckman Coulter (USA). CD3⁺CD4⁺ cells represent T helper cells, CD3+CD8+ cells represent T suppressor cells, and CD4⁺CD25⁺CD127^Dim cells represent T regulatory cells, respectively. Lymphocyte subsets were detected using a flow cytometer (FC500MCL; Beckman Coulter, USA). Hemoglobin A1c was detected using a glycated hemoglobin analyzer (Premier Hb9210, USA).

Chen T., Chen H., Chen P., Zhu L., Mao W, & Yao Y. (2023). High expression of IL6 and decrease in immune cells in COVID-19 patients combined with myocardial injury. Frontiers in Immunology, 14, 1190644.

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Comprehensive T Cell Subset Analysis

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Flow cytometry was carried out in order to analyse the T lymphocyte subsets (T. Chen et al. Citation2020). CD3 + CD45 + cells indicated the total T cells. Various T lymphocyte subsets, containing T helper cells (CD3 + CD4 + ), T suppressor cells (CD3 + CD8 + ), and regulatory T cells (Tregs; CD4 + CD25 + CD127 Dim ) were individually detected with an FC500MCL flow cytometer (Beckman Coulter, Chaska, MN, USA) using corresponding antibodies anti-CD3-PC5, anti-CD4-PE, anti-CD8-ECD, anti-CD4-PC5, anti-CD25-FITC, and anti-CD127-PE (all procured from Beckman Coulter). Briefly, 2 mL venous peripheral blood samples were collected into EDTA tubes for cytometric analysis, 50 μL whole blood per tube was incubated with monoclonal antibodies for 30 min in conditions void of light, and thereafter red blood cells were lysed using a buffer comprising 0.8% NH 4 Cl and 0.1% KHCO 3 (pH 7.1-7.4). Following a rinse with phosphate-buffered saline (PBS), the cells were suspended with 200 µL PBS for detection of T lymphocyte subsets. The percentages of CD3 + T cells, CD3 + CD4 + T cells, CD3 + CD8 + T cells, and Tregs were calculated using the BD Multitest software (BD Biosciences, San Jose, CA, USA).

Ao M., Li P., Sun D., Li X., Xu S, & Hao Y. (2023). Changes in T lymphocyte subsets in peripheral blood of patients with middle-advanced cervical cancer before and after nimotuzumab combined with concurrent chemoradiotherapy. Journal of obstetrics and gynaecology : the journal of the Institute of Obstetrics and Gynaecology, 43(1).

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