Anti rabbit igg bv421

Manufactured by BioLegend

Anti-rabbit IgG BV421 is a secondary antibody conjugated with the BV421 fluorescent dye. It is designed to detect and bind to rabbit primary antibodies in immunoassays.

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3 protocols using anti rabbit igg bv421

Multiparametric Immunofluorescence Staining

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tissue sections were stained with titrated amounts of the following antibodies; Primary: anti-HELIOS (N3C3, GeneTex), anti-PD-1 (goat polyclonal, R&D Systems), anti-BCL-6 (PG-B6p, Dako), anti-IgD (EPR6146, Abcam), ant-CXCL13 (goat polyclonal, ThermoFisher Scientific); Secondary: anti-mouse IgG1 Alexa Fluor 546 and anti-goat IgG Alexa Fluor 546 (Life Technologies) and anti-rabbit IgG BV421 (Biolegend); Primary-conjugated: Ki-67-BV510 (B56, BD Biosciences), CD4-Alexa488 (goat polyclonal, R&D Systems), FOXP3 Alexa647 (206D, Biolegend), JOJO-1 iodide (for nuclear staining, Life Technologies). The CD20-PB (L26) antibody was conjugated in-house.

Amodio D., Cotugno N., Macchiarulo G., Rocca S., Dimopoulos Y., Castrucci M.R., De Vito R., Tucci F.M., McDermott A.B., Narpala S.R., Rossi P., Koup R.A., Palma P, & Petrovas C. (2017). Quantitative multiplexed imaging analysis reveals a strong association between immunogen-specific B cell responses and tonsillar germinal center immune dynamics in children after influenza vaccination. Journal of immunology (Baltimore, Md. : 1950), 200(2), 538-550.

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Immunostaining and Flow Cytometry Protocols

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The following Abs were used for IF and IHC staining: anti-mouse IgA Ab-PE (Abcam, 97013) or fluorescein isothiocyanate (FITC; Abcam, 97234), anti-mouse CD138-PE (clone 281-2, BioLegend), anti-human IgA (DAKO, #IR51061), or CD138 (DAKO, M7228). For flow cytometry, the following Abs against mouse antigens were used: B220–APC-Cy7 (clone RA3-6B2, BioLegend), CD138-PE or BV421 (clone 281-2), GL7-PerCP Cy5.5 (clone GL7, BioLegend), Ki67–PE-Cy7 (clone B56, BD), IgG1-FITC (clone A85-1, BD), IgG2a/c-FITC (clone RMG2a-62, BioLegend), IgG2b-FITC (SouthernBiotech, 1092-02), IgG3-FITC (SouthernBiotech, 1102-02), IgA-biotin (clone RMA-1, BioLegend), CD73-biotin (clone TY/11.8, BioLegend), CD140b-APC (clone APB5, BioLegend), CD105–PE-Cy7 (clone MJ7/18, BioLegend), CD45–PerCP-Cy5.5 (clone 30-F11, BioLegend), and CD31-PE (clone 390, BioLegend); other reagents were also used: rabbit polyclonal anti-SPTBN1 (amino acids 2100 to 2150; Abcam, 72239), normal rabbit IgG (an isotype-matched control for the latter; Sigma-Aldrich, 12-370), anti-rabbit IgG-BV421 (clone Poly4064, BioLegend), streptavidin-BV421 (BioLegend, 405226), or APC (BioLegend, 405207).

Nihei Y., Haniuda K., Higashiyama M., Asami S., Iwasaki H., Fukao Y., Nakayama M., Suzuki H., Kikkawa M., Kazuno S., Miura Y., Suzuki Y, & Kitamura D. (2023). Identification of IgA autoantibodies targeting mesangial cells redefines the pathogenesis of IgA nephropathy. Science Advances, 9(12), eadd6734.

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Phenotypic analysis of CAR T cells

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The following reagents were used for phenotypic analysis of CAR T cells: CD2 APC (Miltenyi), CD3 PerCPCy5.5 (Biolegend), CD4 PE-Vio770 (Miltenyi), CD8 PE or FITC (Biolegend), CD19 BV605 (Biolegend), CD19 PE (Biolegend), CD45RA BV605 (Biolegend), CCR7 APC (Biolegend), CD107a FITC (BD), CD223 APC-eFluor 780 (LAG-3, eBioscience), CD279 BV421 (PD1 Biolegend), CD366 (TIM3 BV711), IFN-gamma APC, TNF-alpha BV421, IL-2 BV605 (Biolegend), Anti-Rabbit Goat F(ab')2 FITC (Jackson Immunoresearch), Anti-Rabbit IgG BV421 (Biolegend), Anti-Mouse IgG PE (Biolegend), Fixable viability dye Life technologies Aqua. Fluorescence minus one (FMO) controls were used to determine expression thresholds where required.

Ghorashian S., Kramer A.M., Onuoha S., Wright G., Bartram J., Richardson R., Albon S.J., Casanovas-Company J., Castro F., Popova B., Villanueva K., Yeung J., Vetharoy W., Guvenel A., Wawrzyniecka P.A., Mekkaoui L., Cheung G.W., Pinner D., Chu J., Lucchini G., Silva J., Ciocarlie O., Lazareva A., Inglott S., Gilmour K.C., Ahsan G., Ferrari M., Manzoor S., Champion K., Brooks T., Lopes A., Hackshaw A., Farzaneh F., Chiesa R., Rao K., Bonney D., Samarasinghe S., Goulden N., Vora A., Veys P., Hough R., Wynn R., Pule M.A, & Amrolia P.J. (2019). Enhanced CAR T cell expansion and prolonged persistence in pediatric patients with ALL treated with a low-affinity CD19 CAR. Nature medicine, 25(9).

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