Anti cd38 fitc

Manufactured by STEMCELL

Sourced in United States, Canada

Anti CD38 FITC is a fluorescently labeled antibody that binds to the CD38 surface antigen. CD38 is expressed on various cell types, including lymphocytes, plasma cells, and myeloid cells. The FITC (fluorescein isothiocyanate) label allows for the detection and analysis of CD38-positive cells using flow cytometry.

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CD38-FITC (8 citations)

4 protocols using anti cd38 fitc

Isolation and Characterization of B Cells

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Frozen lymph node (LN) cell suspensions collected from macaques vaccinated with VLP-RC1–4fill^{12 (link)} were thawed and incubated in FACS buffer (1xPhosphate-buffered saline (PBS), 2% calf serum, 1 mM EDTA) with the following anti human antibodies at 1:200 dilution: anti CD3-APC-eFluor 780 (Invitrogen, 47-0037-41), anti CD14-APC-eFluor 780 (Invitrogen, 47-0149-42), anti CD16-APC-eFluor 780 (Invitrogen, 47-0168-41), anti CD8-APC-eFluor 780 (Invitrogen, 47-0086-42), anti CD20-PE-Cy7 (BD biosciences, 335793), and anti CD38 FITC (Stem Cell, 60131FI) and the LIVE/DEAD marker (Biolegend, 77184) at 1:400 dilution. Avi-tagged and biotinylated baits conjugated to fluorophore labeled streptavidin were added to the antibody mixture and incubated with the LN cells for 30 min. Single B cells were sorted into individual wells of a 96-well plates containing 5 μl of TCL lysis buffer (Qiagen, 1031576) and 1% beta-mercaptoethanol per well using a FACS Aria III (Becton Dickinson). The cell lysates were stored at −80°C or immediately used for subsequent mRNA purification.
To interrogate the B cell receptor specificity of the retrogenic B cells, the transduced B cells were incubated with Avi-tagged-biotinylated RC1 in complex with fluorophore labeled streptavidin for 30 min on ice. Alternatively, cells were incubated with fluorophore labeled streptavidin in the absence of RC1 for 30 min on ice.

Wang Z., Merkenschlager J., Chen S.T., Oliveira T.Y., Ramos V., Gordon K.M., Yao K.H., Jankovic M., Gazumyan A., Nussenzweig M, & Escolano A. (2019). Isolation of single HIV-1 Envelope specific B cells and antibody cloning from immunized rhesus macaques. Journal of immunological methods.

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Comprehensive Immune Cell Profiling

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The following mAbs were used for flow cytometry: anti-CD3 PerCP, anti-CD4 FITC, anti-CD4 PerCP, anti-CD4 PE-Cy7, anti-CD8 PErCP, anti-CD8 APC-Cy7, anti-CD28 FITC, anti-CD95 APC, anti-CD195 (anti-CCR5) PE, anti-Ki67 PE, anti-HLA-DR APC, and anti-HLA-DR FITC (BD Biosciences). Additionally, annexin V PE was purchased from BD Biosciences, anti-CD8 APC and anti-CD8 PE were obtained from Beckman Coulter (CA, USA), and anti-CD38 FITC was purchased from StemCell Technologies (Vancouver, Canada). Flow cytometry data were acquired on a FACSAria II (BD Biosciences). All data were analyzed using FlowJo software (Tree Star, Ashland, OR, USA).

Zhan X.Y., Wang N., Liu G., Qin L., Xu W., Zhao S., Qin L, & Chen X. (2014). Plasmodium infection reduces the volume of the viral reservoir in SIV-infected rhesus macaques receiving antiretroviral therapy. Retrovirology, 11, 112.

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Comprehensive Multiparametric Flow Cytometry Analysis of CD4+ T Cell Subsets and Activation States

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For CD4 T cell subsets sort, PBMC were stained with fluorochrome-labelled mAbs anti-CD28-CY5PE, anti-CCR5-PE, anti-CD3-CY7APC, anti-CD4-BV605, anti-CD8-Pacific blue (BD Biosciences) and anti-CD95-BV785 (in house conjugated, BD Biosciences). LN cells were stained with anti-CD28-CY5PE, anti-CD3-CY7APC, anti-CD4-BV605 (BD Biosciences), anti-CD8-BV570, anti-CXCR5-PE (eBioscience) and anti-CD95-BV785 (in house conjugated, BD Biosciences). PBMC and LN CD4 subsets of interest were sorted and lyzed in proteinase K (100ug mL^-1, Sigma Aldrich) for SIV gag qPCR. For assessment of T cell activation and exhaustion, cryopreserved PBMC were thawed and stained with fluorochrome-labelled mAbs anti-CD38-FITC (Stem Cell), anti-Ki67-CY7PE, anti-CD28-CY5PE, anti-CD3-CY7APC (BD Biosciences), anti-HLA-DR-TRPE (Life Technologies), anti-PD-1-BV711, anti-CD95-BV785 (Biolegend), anti-TIGIT-APC (ThermoFisher) and anti-LAG3-PE (R&D). All samples were stained with Aqua LIVE/DEAD Fixable Dead Cell Stain.

Nganou-Makamdop K., Billingsley J.M., Yaffe Z., O’Connor G., Tharp G.K., Ransier A., Laboune F., Matus-Nicodemos R., Lerner A., Gharu L., Robertson J.M., Ford M.L., Schlapschy M., Kuhn N., Lensch A., Lifson J., Nason M., Skerra A., Schreiber G., Bosinger S.E, & Douek D.C. (2018). Type I IFN signaling blockade by a PASylated antagonist during chronic SIV infection suppresses specific inflammatory pathways but does not alter T cell activation or virus replication. PLoS Pathogens, 14(8), e1007246.

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Phenotypic Analysis of T-cells by Flow Cytometry

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Flow cytometry analysis of T-cell phenotype was performed according to the protocols as previously described. [21] [22] [23] Monoclonal antibodies used in this study including anti-CD3-BV605, anti-CD45-PE, anti-CD4-BV421, anti-CD8-APC-R700 anti-CD95-DX2, anti-CD28-CD28.2, anti-HLA-DR-PE-Cy7, anti-CD69-APC and anti-CCR5-PE, were purchased from BD Biosciences (San Jose, CA, USA). Anti-CD38-FITC was obtained from StemCell Technologies (Vancouver, BC, Canada). Anti-CD25-APC was from Biolegend (San Diego, CA, USA), and anti-CD127-PE was a product of Invitrogen (Carlsbad, CA, USA). CD4 + T cell differentiation was identi ed in terms of CD28 and CD95 expression, as CD28 + CD95 + CD4 + T cell de ned as central memory CD4 + T cell (CD4 + Tcm) and CD28 + CD95 -CD4 + T cell as effector memory CD4+ T cell (CD4 + T EM ). CD28 + CD95 + CD8 + T cell was de ned as CD8 + T CM cell whereas CD28 + CD95 -CD8 + T cell was CD8 + T EM cell. In addition, expression of CD25 and CD127 were measured to evaluate regulatory T Cells (Tregs). Activation markers HLA-DR, CD38 and CD69 were measured on CD4 + and CD8 + T cells, and CD4 + CCR5 + T cells were de ned as SIV-infected cells. All dates were acquired and analyzed on a ow cytometer (FACSCanto; Becton Dickinson).

Wang Q., Zeng Q., Ren Z., Li Y., Wang R., Feng Y., Ye J., Song X., Qin S., Zhou P., Zhu Y., Feng L., Li Z., Qi H., Li Q., Jin F., & Wang Y. (2021). Traditional Chinese Medicine TN-01 Reconstitutes T Cells in SIV–infected Rhesus Monkeys.

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