Anti cd38 apc cy7

Manufactured by BioLegend

Sourced in United States

Anti-CD38 APC-Cy7 is a fluorochrome-conjugated monoclonal antibody that specifically binds to the CD38 antigen. CD38 is a cell surface glycoprotein expressed on various cell types, including plasma cells, activated T and B cells, and natural killer cells. This antibody can be used for the identification and enumeration of CD38-positive cells in flow cytometry applications.

Automatically generated - may contain errors

Lab products found in correlation

6 protocols using anti cd38 apc cy7

HLA Class II Tetramer Staining Protocol

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Direct HLA class II tetramer staining was performed essentially as previously described (11 , 12 (link)), except that cells were labeled with PE and PE-CF594 labeled tetramers. Briefly, 30×10⁶ PBMCs were re-suspended in 200 µL of T cell media, incubated with 50 nM dasatinib for 10 minutes at 37°C, and stained with PE-DRB4/PPI 9–28 tetramer and PE-CF594-DR0401/PPI 76–90 (with an Lys Ser substitution at position 88, as described (13 (link))) at room temperature for 100 minutes. Cells were washed, incubated with anti-PE magnetic beads (Miltenyi) for 20 minutes at 4°C and enriched with a magnetic column, retaining 1% of the cells as a non-enriched sample. The enriched and pre-column samples were labeled with anti-CD4 PerCP-Cy5.5 (Biolegend), anti-CD14 and anti-CD19 FITC (both eBioscience), ant-CD45RA AF700 (BD Biosciences), anti-CXCR3 APC (BD Biosciences), anti-CD38 APC-Cy7 (Biolegend), and SYTOX Green (ThermoFisher) as a viability indicator for 15 minutes at 4°C and analyzed on an LSR II (BD Biosciences), gating on Viable CD4⁺ cells and excluding events that were positive for more than one tetramer color. Frequencies of tetramer positive cells were calculated as previously described (12 (link)).

James E.A., Gillette L., Durinovic-Bello I., Speake C., Bondinas G.P., Moustakas A.K., Greenbaum C.J., Papadopoulos G.K, & Kwok W.W. (2018). DRB4 has a distinct motif and presents a proinsulin epitope that is recognized in subjects with type 1 diabetes. Journal of immunology (Baltimore, Md. : 1950), 201(12), 3524-3533.

+ Open protocol

+ Expand

Antigen-experienced B Cell Sorting

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cryopreserved PBMCs were rapidly thawed in a 37°C water bath for 1 min and then washed with 10 mL cold PBS. After centrifugation, the supernatant was removed, and the PBMCs were resuspended in PBS supplemented with 2% FBS (Cytiva). PBMCs were incubated with Fc-blocker (BioLegend) followed by staining with an antibody cocktail containing anti CD19-AF488 (BioLegend), anti CD3-BV421 (BioLegend), anti CD27-PE (BioLegend), and anti CD38-APC.Cy7 (BioLegend) for 30 min together with fluorescence-labeled antigen probes. Additionally, four different Total-Seq C anti-human hashtag antibodies (BioLegend) were used to label the donor. After three washes, cells from different donors were combined in FACS buffer (2% FBS in PBS) and sorted using BD Aria II Cell Sorter, with the gating strategy described in Fig. S1B. Antigen-experienced B cells (CD19⁺CD27⁺) were sorted, and double negative controls were utilized to gate the toxin- and non-binding cells: unlabeled toxin and the non-conjugated anti-His APC (BioLegend) for DT and TT, or non-conjugated Alexa Fluor 647 (Invitrogen) for PT.

Saputri D.S., Ismanto H.S., Nugraha D.K., Xu Z., Horiguchi Y., Sakakibara S, & Standley D.M. (2023). Deciphering the antigen specificities of antibodies by clustering their complementarity determining region sequences. mSystems, 8(6), e00722-23.

+ Open protocol

+ Expand

Protocol for Immunophenotyping of Human B Cell Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols

For cell sorting, cell suspension were incubated at 4°C with Fc-blocking reagent (Miltenyi Biotec) and stained for 30min with the following monoclonal antibodies: anti-CD45 AF700 (clone: HI30), anti-CD19 PE-Cy7 (clone: HIB19), anti-CD38APC-Cy7 (clone: HIT2), anti-CD10 PE (clone: HI10a), anti-IgM BV605 (clone: MHM-88) (all from Biolegend), anti-CD27 PerCpCy5.5 (clone: M-T271) (BD Biosciences), and anti-IgD FITC (Southern). CD45⁺CD19⁺CD38^dullCD10⁻IgD²⁺ IgM⁺CD27⁻ naive B cells, CD45⁺CD19⁺CD38^dullCD10⁻IgD⁺IgM²⁺CD27⁺ MZ B cells, CD45⁺CD19⁺CD38^intCD10⁺IgD-IgM⁺CD27⁺ GC-M, CD45⁺CD19⁺CD38^dullCD10⁻IgD⁻IgM⁺ CD27⁺ ME-M B cells, CD45⁺CD19⁺ CD38^dullCD10⁻ IgD⁻IgM⁻CD27⁺ ME-SW B cells, CD45⁺CD19⁺CD38^intCD10⁺IgD⁻CD27⁺ GC B cells, CD45⁺CD19⁺CD38²⁺CD10⁻IgD⁻IgM⁺CD27⁺ PC-M and CD45⁺CD19⁺CD38²⁺CD10⁻ IgD⁻IgM⁺CD27⁺ switched PC were sorted with a FACSAria II (BD Biosciences) after exclusion of dead cells through DAPI staining. For sorting of FCRL4⁺ and FCRL4^- ME-M, anti-FCRL4 APC (clone: 413D12) was added to the ‘cocktails’. The purity of cells sorted this way was consistently > 95%.

Magri G., Comerma L., Pybus M., Sintes J., Lligé D., Segura-Garzón D., Bascones S., Yeste A., Grasset E.K., Gutzeit C., Uzzan M., Ramanujam M., van Zelm M.C., Albero-González R., Vazquez I., Iglesias M., Serrano S., Márquez L., Mercade E., Mehandru S, & Cerutti A. (2017). Human Secretory IgM Emerges from Plasma Cells Clonally Related to Gut Memory B Cells and Targets Highly Diverse Commensals. Immunity, 47(1), 118-134.e8.

+ Open protocol

+ Expand

Multiparametric Flow Cytometry of SARS-CoV-2 Spike Antibodies

Check if the same lab product or an alternative is used in the 5 most similar protocols

Flow cytometry of cryopreserved PBMCs was performed on a BD FACS Melody as previously described. SARS-CoV-2 Wuhan-1 Spike was pre-complexed with the streptavidin fluorophore (Alexa-488) at a 4:1 molar ratio prior to addition to cells. PBMCs were incubated with Fc block for 15 minutes at 4°C. PBMCs were stained with anti-CD3-BV510 (Biolegend), anti-CD19-PerCP-Cy5.5 (Biolegend), anti-IgD-Pacific Blue (Biolegend), anti-CD27-BV785 (Biolegend), anti-CD38-APC-Cy7 (Biolegend), anti-CD20-APC (Biolegend), anti-IgG-PE-Cy7 (BD Biosciences), anti-IgM-PE (Biolegend) and Spike-Alexa Fluor 488 for 1 hour at 4°C. PBMCs were washed with PBS and stained with live/dead for 1 hour at 4°C.

Graham C., Lechmere T., Rehman A., Seow J., Kurshan A., Huettner I., Maguire T.J., Tam J.C., Cox D., Ward C., Racz M., Waters A., Mant C., Malim M.H., Fox J, & Doores K.J. (2022). The effect of Omicron breakthrough infection and extended BNT162b2 booster dosing on neutralization breadth against SARS-CoV-2 variants of concern. PLoS Pathogens, 18(10), e1010882.

+ Open protocol

+ Expand

Multiparameter Flow Cytometry Analysis of Immune Cell Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols

Samples were blocked with mouse FcR Blocking Reagent (MiltenyiBiotec, USA), and then stained with the indicated antibodies. The antibodies included APC/Cy7 anti-interferon (IFN)γ, APC/Cy7 anti-CD38, BV421 anti-CCR7, BV510 anti-CXCR3, PerCP/Cy5.5 anti-CXCR5, biotinylated anti-CXCR5 from Biolegend (USA); AF488 anti-CD4, eFluor506 anti-IL17A, PE anti-IL21, PE-Cy7 anti-CD44, PE-Cy7 anti-FoxP3 from eBiosciences (USA); FITC anti-CD69, PE anti-CD25, PerCP anti-CD3e, PE-Cy7 anti-IL4 from BD Biosciences (USA); biotinylated anti-PD-1 from R&D systems (USA) and streptavidin-APC from Jackson Immunoresearch (USA). When necessary, red blood cell lysis was performed using BD Pharm Lyse lysing buffer (BD Biosciences). For intracellular staining, CytoFix/Perm kit (BD Biosciences) or FoxP3/Transcription Factor Staining Buffer Set (eBiosciences) were used according to the manufacturer’s instructions. Cytometric data were collected using an FACS Canto II (BD Biosciences) and analyzed using FlowJo software (BD Biosciences). Gating strategy is shown in online supplemental figure 4.

Sánchez-Alonso S., Setti-Jerez G., Arroyo M., Hernández T., Martos M.I., Sánchez-Torres J.M., Colomer R., Ramiro A.R, & Alfranca A. (2020). A new role for circulating T follicular helper cells in humoral response to anti-PD-1 therapy. Journal for Immunotherapy of Cancer, 8(2), e001187.

+ Open protocol

+ Expand

Multiparametric Flow Cytometry Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

All flow cytometric analyses were performed using a BD FACS Fortessa (BD Biosciences, Franklin Lakes, NJ, USA). To ensure comparable mean fluorescence intensities (MFIs) over time of the analyses, Cytometer Setup and Tracking beads (CST beads, BD Biosciences, Franklin Lakes, NJ, USA) and Rainbow Calibration Particles (BD Biosciences, Franklin Lakes, NJ, USA) were used. For flow cytometric analysis, the following fluorochrome-labeled antibodies were used: BUV737 anti-CD11c (BD, clone B-ly6), BUV395 anti-CD14 (BD, clone M5E2), BUV395 anti-CD3 (BD, clone UCHT1), BV786 anti-CD27 (BD, clone L128), BV711 anti-CD19 (BD, clone SJ25C1), BV605 anti-CD24 (BD, clone ML5), BV510 anti-CD10 (BD, clone HI10A), BV421 anti-CXCR5 (BD, clone RF8B2), PE-CF594 anti-IgD (Biolegend, San Diego, CA, USA, clone IA6-2), APC-Cy7 anti-CD38 (Biolegend, clone HIT2), PE-Cy7 anti-IgG (BD, clone G18-145), anti-IgA-Biotin (BD, clone G20-359), BV650 anti-IgM (BD, clone MHM-88), FITC anti-TNFα (Biolegend, clone Mab11), BV650 anti-IFNγ (BD, clone 4S.B3), BV786 anti-CD40L (Biolegend, clone 24-31), PE-CF594 anti-CD137 (Biolegend, clone 4B4-1). Numbers of absolute B and T cells were measured with Trucount (BD) and samples were processed according to the manufacturer’s instruction.

Stefanski A.L., Rincon-Arevalo H., Schrezenmeier E., Karberg K., Szelinski F., Ritter J., Chen Y., Meisel C., Jahrsdörfer B., Ludwig C., Schrezenmeier H., Lino A.C, & Dörner T. (2022). Persistent but atypical germinal center reaction among 3rd SARS-CoV-2 vaccination after rituximab exposure. Frontiers in Immunology, 13, 943476.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!