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Cd38 apc clone hit2

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CD38-APC (clone HIT2) is a fluorochrome-conjugated monoclonal antibody that binds to the CD38 antigen. CD38 is a transmembrane glycoprotein expressed on various cell types, including lymphocytes, plasma cells, and some hematopoietic progenitor cells. This product is intended for use in flow cytometry applications.

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Lab products found in correlation

Cd26 pe clone m a261, by BD (2 mentions) Anti il6 antibody clone 6708, by R&D Systems (2 mentions) Cd19 bv421 clone hib19, by BD (2 mentions) Cd123 fitc clone 7g3, by BD (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) Hepes, by Thermo Fisher Scientific (2 mentions) β mercaptoethanol, by Merck Group (2 mentions) Cd45 v500, by BD (1 mentions) Cd3 qd655, by Thermo Fisher Scientific (1 mentions) Hla dr fitc clone g46 6, by BD (1 mentions) Cd19 pacific blue clone sj25 c1, by Thermo Fisher Scientific (1 mentions) Cd8 pe cy7, by BD (1 mentions) Cd8 pe cy7 clone sk1, by BioLegend (1 mentions) Hla dr fitc clone l243, by BioLegend (1 mentions) Cd4 bv421, by BioLegend (1 mentions) Canto 2 instrument, by BD (1 mentions) Cd31 fitc clone wm59, by BD (1 mentions) Cd14 pe clone m5e2, by BD (1 mentions) Cd34 fitc clone 581, by BD (1 mentions) Macsquant analyzer, by Miltenyi Biotec (1 mentions)

Spelling variants of this product

CD38-APC (42 citations) CD38 (HIT2, (8 citations) CD38 APC (HIT2, (5 citations)

7 protocols using cd38 apc clone hit2

1

Multiparametric Flow Cytometry of CD34+/CD38-/CD26+ Cells

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Three milliliters of EDTA PB samples (all 211 subjects) or 1 mL of EDTA BM sample (only 84 subjects) were centrally analyzed at Flow‐cytometry lab in Siena within 24 h of collection, since in preliminary experiments it was shown that within 24 h cell viability was superior to 80%. BM and PB leukocytes and red blood cells count was performed using a Unicell DxH 800 Coulter (Beckman Coulter, Brea, CA). In all PB and BM samples, the presence of CD34+/CD38−/CD26+ population was evaluated by multiparametric flow cytometry analysis using a four‐color staining standardized protocol with lyse stain wash procedure. Red cells’ lysis was performed with BD Pharm Lyse™ ammonium chloride (Ref 555899, BD Biosciences, San Jose, CA), 1:10 diluted in deionized water, using BD FACS™ Lyse Wash Assistant (LWA) instrument (BD Biosciences, San Jose, CA). After lysis, 2.0 × 106 leucocytes per mL were incubated with a custom‐made lyophilized pre‐titrated antibody mixture test and control tube (Ref 625183, BD Biosciences, San Jose, CA), containing CD34‐FITC (clone 581), CD26‐PE (clone M‐A261), CD38‐APC (clone HIT2), and CD45‐V500 (clone 2D1) for tube test and CD34‐FITC (clone 581), anti IGg1 PE, CD38‐APC (clone HIT2), and CD45‐V500 (clone 2D1) for control tube.
Raspadori D., Pacelli P., Sicuranza A., Abruzzese E., Iurlo A., Cattaneo D., Gozzini A., Galimberti S., Baratè C., Pregno P., Nicolosi M., Sorà F., Annunziata M., Luciano L., Caocci G., Moretti S., Sgherza N., Fozza C., Russo S., Usala E., Liberati M.A., Ciofini S., Trawinska M.M., Gozzetti A, & Bocchia M. (2019). Flow Cytometry Assessment of CD26+ Leukemic Stem Cells in Peripheral Blood: A Simple and Rapid New Diagnostic Tool for Chronic Myeloid Leukemia. Cytometry. Part B, Clinical Cytometry, 96(4), 294-299.
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2

Multicolor Flow Cytometry Panel for T Cell Activation

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Fresh EDTA whole blood samples (500 μL) were stained with antibodies CD3-QD655 (clone S4.1, Invitrogen), CD4-QD605 (clone S3.5, Invitrogen), CD8-PECy7 (clone 3B5, BD Biosciences), HLA-DR-FITC (clone G46-6, BD Biosciences), and CD38-APC (clone HIT2, BD Biosciences). CD14-Pacific blue (clone TÜK4) and CD19-Pacific blue (clone SJ25-C1) from Invitrogen were used to exclude monocytes and B cells. Stained cells were acquired on an 18-color LSR II flow cytometer (BD Immunocytometry Systems) and analyzed using FlowJo software. The gating strategy (Supplementary Figure 2) was to gate on lymphocytes in the scatterplot, then singlets, then live CD3+ and CD14– and CD19– cells. From the CD3+ cell population, CD4+ and CD8+ cell populations were selected. Activation was measured as percentage of CD38+HLA-DR+CD8+ or CD38+HLA-DR+CD4+ T cells.
Kyosiimire-Lugemwa J., Anywaine Z., Abaasa A., Levin J., Gombe B., Musinguzi K., Kaleebu P., Grosskurth H., Munderi P, & Pala P. (2019). Effect of Stopping Cotrimoxazole Preventive Therapy on Microbial Translocation and Inflammatory Markers Among Human Immunodeficiency Virus–Infected Ugandan Adults on Antiretroviral Therapy: The COSTOP Trial Immunology Substudy. The Journal of Infectious Diseases, 222(3), 381-390.
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3

T Cell Activation Analysis in PBMC Samples

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PBMC samples were stained with the following antibodies for T cell activation analysis: CD3-BV510 (clone OKT3), CD4-BV421 (clone OKT4), CD8-PE-Cy7 (clone SK1), HLA-DR-FITC (clone L243; Biolegend, San Diego, CA, USA); CD38-APC (clone HIT2; BD Biosciences, San Diego, CA, USA). The BD Canto II instrument (BD Biosciences, San Diego, CA, USA) was used for data collection, and the data was analyzed using FlowJo software V10 (Tree star Inc., Ashland, OR, USA).
Fan X., Song J.W., Wang S.Y., Cao W.J., Wang X.W., Zhou M.J., Yang T., Zhou C.B., Hou J., Zhang J.Y., Meng F.P., Shi M., Wang F.S, & Zhang C. (2021). Changes of Damage Associated Molecular Patterns in COVID-19 Patients. Infectious Diseases & Immunity, 1(1), 20-27.
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4

In Vitro B Cell Differentiation Assay

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B cells and pDCs were resuspended in RPMI 1640 medium supplemented with 10% FBS, 1x penicillin/streptomycin, 2mM L-glutamine, 5mM HEPES (all from Life Technologies) and 55 μM β-mercaptoethanol (Sigma) and transferred into U-bottom 96-well plates. 7.5 x 104 B cells and 3.75 x 103 pDCs were added to each well, along with 1.5 nM mega CD40L (Enzo Biosciences). ICs generated by combining 2.5 μg/mL of IgED with 1 μg/mL of CG50 plasmid DNA were added to the co-cultures. Cells were cultured for 7 days at 37°C and 5% CO2, washed and stained with CD123-FITC (clone 7G3), CD19-BV421 (clone HIB19), CD27-PerCPCy5.5 (clone M-T271), IgD-PE (clone IA6-2) and CD38-APC (clone HIT2), all from BD Biosciences. For flow cytometry assessment of B cell and plasma cell numbers, co-cultured cells were acquired for a fixed amount of time. B cells were defined as CD123− CD19+ and plasma cells were defined as CD123−CD19+ CD27hi CD38hi cells. Supernatants were recovered at Day 7 for indirect quantification of IgM by ELISA as described previously48 (link). Anti-human IFN-α receptor (IFNAR) antibody (MedImmune LLC)49 (link) anti-IL6 antibody (clone 6708, R&D Systems), or an anti-IL6 antibody (MedImmune LLC)50 (link) were used at final concentrations of 10 μg/mL.
Henault J., Riggs J.M., Karnell J.L., Liarski V.M., Li J., Shirinian L., Xu L., Casey K.A., Smith M.A., Khatry D.B., Izhak L., Clarke L., Herbst R., Ettinger R., Petri M., Clark M.R., Mustelin T., Kolbeck R, & Sanjuan M.A. (2015). Self-reactive IgE exacerbates interferon responses associated with autoimmunity. Nature immunology, 17(2), 196-203.
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5

Immunophenotyping of Multiple Myeloma Cells

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The immunophenotype of primary BM CD138+ cells, HMCLs and microenvironment cells was analyzed with the following mAbs:
CD38-APC (clone HIT2, code n. 560677, BD)
CD31-FITC (clone WM59, code n. 555445, BD)
CD39-APC (clone eBioA1, code n. 17-0399-41, eBioscence; San Diego, CA)
CD73-APC (clone CB73), produced in the Lab of one of the authors (FM) and FITC-conjugated by AcZon (Bologna, Italy)
CD203a-FITC (clone 3E8, kindly provided by J. Goding)
CD14-PE (clone M5E2, code n. 555398, BD).
Costa F., Toscani D., Chillemi A., Quarona V., Bolzoni M., Marchica V., Vescovini R., Mancini C., Martella E., Campanini N., Schifano C., Bonomini S., Accardi F., Horenstein A.L., Aversa F., Malavasi F, & Giuliani N. (2017). Expression of CD38 in myeloma bone niche: A rational basis for the use of anti-CD38 immunotherapy to inhibit osteoclast formation. Oncotarget, 8(34), 56598-56611.
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6

In Vitro B Cell Differentiation Assay

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B cells and pDCs were resuspended in RPMI 1640 medium supplemented with 10% FBS, 1x penicillin/streptomycin, 2mM L-glutamine, 5mM HEPES (all from Life Technologies) and 55 μM β-mercaptoethanol (Sigma) and transferred into U-bottom 96-well plates. 7.5 x 104 B cells and 3.75 x 103 pDCs were added to each well, along with 1.5 nM mega CD40L (Enzo Biosciences). ICs generated by combining 2.5 μg/mL of IgED with 1 μg/mL of CG50 plasmid DNA were added to the co-cultures. Cells were cultured for 7 days at 37°C and 5% CO2, washed and stained with CD123-FITC (clone 7G3), CD19-BV421 (clone HIB19), CD27-PerCPCy5.5 (clone M-T271), IgD-PE (clone IA6-2) and CD38-APC (clone HIT2), all from BD Biosciences. For flow cytometry assessment of B cell and plasma cell numbers, co-cultured cells were acquired for a fixed amount of time. B cells were defined as CD123− CD19+ and plasma cells were defined as CD123−CD19+ CD27hi CD38hi cells. Supernatants were recovered at Day 7 for indirect quantification of IgM by ELISA as described previously48 (link). Anti-human IFN-α receptor (IFNAR) antibody (MedImmune LLC)49 (link) anti-IL6 antibody (clone 6708, R&D Systems), or an anti-IL6 antibody (MedImmune LLC)50 (link) were used at final concentrations of 10 μg/mL.
Henault J., Riggs J.M., Karnell J.L., Liarski V.M., Li J., Shirinian L., Xu L., Casey K.A., Smith M.A., Khatry D.B., Izhak L., Clarke L., Herbst R., Ettinger R., Petri M., Clark M.R., Mustelin T., Kolbeck R, & Sanjuan M.A. (2015). Self-reactive IgE exacerbates interferon responses associated with autoimmunity. Nature immunology, 17(2), 196-203.
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7

CD26 Expression in Bone Marrow Progenitors

Cited 1 time
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For each patient 2 mL of bone marrow samples collected in EDTA tubes were analyzed. CD26 expression was assessed by multiparametric flow cytometry, analyzing the CD45+/CD34+/CD38− population using a four-color protocol, as previously reported by our group (15 (link)). Briefly, 2.0 x 106 leucocytes were incubated with BD Pharmigen CD45-V500 (clone 2D1), CD34-FITC (clone 581), CD38-APC (clone HIT2), CD26-PE (clone M-A261), and proper negative controls. After washing, acquisition and analysis of at least 1.0 × 106 of CD45+ cells were performed by using a MACSQuant Analyzer (Miltenyi, Gemany), equipped with 3 lasers and 8 fluorescent channels available. The absolute number of CD26+ cells was calculated by multiplying the number of white cells/μL automatically counted for the proportion of CD34+/CD38−/CD26+ on CD45+ cells using the MACSQuantify Software (Miltenyi).
Galimberti S., Grassi S., Baratè C., Guerrini F., Ciabatti E., Perutelli F., Ricci F., Del Genio G., Montali M., Barachini S., Giuliani C., Ferreri M.I., Valetto A., Abruzzese E., Ippolito C., Iurlo A., Bocchia M., Sicuranza A., Martino B., Iovino L., Buda G., Salehzadeh S., Petrini M., Di Paolo A, & Mattii L. (2018). The Polycomb BMI1 Protein Is Co-expressed With CD26+ in Leukemic Stem Cells of Chronic Myeloid Leukemia. Frontiers in Oncology, 8, 555.
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