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Igd ia6 2

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IgD (IA6-2) is a mouse monoclonal antibody that recognizes the human IgD protein. It is a laboratory reagent used for the detection and study of IgD in various applications.

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Lab products found in correlation

Mitotracker orange, by Thermo Fisher Scientific (3 mentions) Ficoll paque plus gradient, by GE Healthcare (1 mentions) Cd27 m t271, by BioLegend (1 mentions) Foxp3 pch101, by Thermo Fisher Scientific (1 mentions) Facsaria cell sorter, by BD (1 mentions) Ramos cells, by American Type Culture Collection (1 mentions) 2 nbdg, by Thermo Fisher Scientific (1 mentions) Cd27 m t271, by BD (1 mentions) Cd3 hit3a, by BioLegend (1 mentions) 2 nbdg, by American Type Culture Collection (1 mentions) Cd10 hi10a, by BioLegend (1 mentions) Cellrox, by Thermo Fisher Scientific (1 mentions) Igm pe, by Thermo Fisher Scientific (1 mentions) Anti igd apc, by Thermo Fisher Scientific (1 mentions) Anti cd19 pecy7, by Thermo Fisher Scientific (1 mentions) Anti cd20 apc, by Thermo Fisher Scientific (1 mentions) Anti igm apc, by Thermo Fisher Scientific (1 mentions)

3 protocols using igd ia6 2

1

Immunophenotyping of PBMC Subsets

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Peripheral blood mononuclear cells (PBMCs) were isolated on a Ficoll-Paque PLUS gradient (GE Healthcare; IL, USA). Anti-human antibodies for surface and intracellular makers with isotype-matched controls were used for staining CD3 (UCHT1), CD4 (OKT4), CD8 (RPA-T8), CD45RA (HI100), CD45RO (UCHL1), CCR7 (G043H7), CD31 (WM59), CD16 (3G8), CD56 (5.1H11), CD127/IL-7Ra (A019D5), CD25 (BC96), CTLA-4 (L3D10), Helios (22F6), CD19 (HIB19), CD27 (M-T271), and IgD (IA6-2), all from Biolegend (CA, USA), and FOXP3 (PCH101) from eBioscience (CA, USA). Cells were analyzed on a BD FACS Aria cell sorter (BD Biosciences, NJ, USA), and analyzed with FlowJo software (Tree Star Inc.; OR, USA).
Mansour R., Bsat Y.E., Fadel A., El-Orfali Y., Noun D., Tarek N., Kabbara N., Abboud M, & Massaad M.J. (2022). Diagnosis and Treatment of a Patient With Severe Combined Immunodeficiency Due to a Novel Homozygous Mutation in the IL-7Rα Chain. Frontiers in Immunology, 13, 867837.
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2

Multiparametric Analysis of Immune Cell Function

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Fresh healthy donor human PBMCs were incubated with CellROX (Thermo Fisher Scientific), NAO (Thermo Fisher Scientific), or 2-NBDG (Carbosynth) according to the manufacturer’s specifications. For all three conditions, cells were stained during the efflux incubation for MitoTracker and otherwise prepared for flow cytometry using the protocol detailed above with a few exceptions. 2-NBDG and NAO panels required substitution of MitoTracker Orange (Thermo Fisher Scientific). The fluorochrome-conjugated primary antibody panel was as follows: CD3 (HIT3a, BioLegend), CD19 (SJ25C1, BioLegend), CD27 (M-T271, BD Biosciences), IgD (IA6–2, BioLegend), and CD10 (HI10a, BioLegend). Compensation for the functional dyes required the use of Ramos cells (ATCC), incubated with CellROX, 2-NBDG, or NAO, respectively.
Farmer J.R., Allard-Chamard H., Sun N., Ahmad M., Bertocchi A., Mahajan V.S., Aicher T., Arnold J., Benson M.D., Morningstar J., Barmettler S., Yuen G., Murphy S.J., Walter J.E., Ghebremichael M., Shalek A.K., Batista F., Gerszten R, & Pillai S. (2019). Induction of metabolic quiescence defines the transitional to follicular B cell switch. Science signaling, 12(604), eaaw5573.
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3

FACS and PLA Analysis of Mouse and Human B Cells

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For FACS analysis of mouse spleen B cells or transfected TKO cells, following fluorophore- conjugated anti-mouse antibodies were used: Anti-CD45R-PerCP-Cy5.5 (RA3-6B2), Anti-IgM-APC, IgM-PE, IgM-FITC (II/41; all eBioscience, Frankfurt, Germany), Anti-CD20-APC, Anti-CD19-PE-Cy7, anti-IgD-AF647, Anti-IgD-APC, Anti-IgD-PE (11-26c.2a; all eBioscience).
For PLA probes against specific targets, the following unlabelled antibodies were used: IgD (11-26c.2a, SouthernBiotech, Birmingham, AL), IgD (AMS9.1; Santa Cruz Biotechnology, Dallas, TX), IgM (R33.24.12, in house hybridoma culture), IgM (rabbit anti-mouse µHC; Rockland Immunochemicals, Gilbertsville, PA), IgM (1B4B1; SouthernBiotech), Lambda light chain (JC5-1; SouthernBiotech), Kappa light chain (187.1; SouthernBiotech), CD19 (6D5; AbD Serotec, Düsseldorf, Germany) and CD20 (AISB12; eBioscience). Igα (HMK7/A9; abcam, Cambridge, UK), Syk (Syk-01; BioLegend, San Diego, CA).
For PLA probes against human BCR, the following unlabelled antibodies were used: IgD (IA6-2; BioLegend), IgD (IADB6) and IgM (SA-DA4) from Acris Antibodies (Herford, Germany), and IgM (Fc5u) from Genway Biotech (San Diego, CA).
Kläsener K., Maity P.C., Hobeika E., Yang J, & Reth M. (2014). B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. eLife, 3, e02069.
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