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Irf8 percp efluor710

Manufactured by Thermo Fisher Scientific
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The Irf8 PerCP-eFluor710 is a laboratory instrument used for the detection and analysis of Irf8 (Interferon Regulatory Factor 8) protein expression in biological samples. This product is designed to provide a reliable and efficient method for researchers to study the role of Irf8 in various cellular processes and disease states.

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3 protocols using irf8 percp efluor710

1

Comprehensive B Cell Immunophenotyping

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B cell subsets were identified with CD22.2-FITC, CD138-PE, CD45.2-PE, CD45.1-APC (BD Biosciences), CD45R/B220-eFluor450, CD44-eFluor780 (eBioscience). RF B cells were detected with biotinylated-4G7 in combination with streptavidin-PerCP-Cy5.5. IRF-4 was detected using an IRF-4 antibody (clone M-17, Santa Cruz) and anti-goat IgG Alexa Fluor 647 (Jackson ImmunoResearch). IRF4-PE and IRF8-PercP-efluor710 (eBioscience) were used to co-stain IRF8 and IRF4. B cell proliferation was assessed by CFSE dilution (Life Technologies) (15 ) or VPD450 dilution (BD). Dead cells were distinguished with TO-PRO-3 (Life Technologies). To analyze TLR7 expression levels, unstimulated purified B cells, or B cells stimulated for 24 hr, were fixed and permeabilized using the Foxp3 Fix/Perm kit (ebioscience). TLR7 protein was detected using a biotinylated mouse TLR7 specific mAb, A94 (25 (link)), in combination with SA-PE. Flow cytometric analysis was carried out using a BD LSR II with Diva Software (BD) and analysis was conducted with FlowJo software (Tree Star).
Nündel K., Green N.M., Shaffer A.L., Moody K.S., Busto P., Eilat D., Miyake K., Oropallo M.A., Cancro M.P, & Marshak-Rothstein A. (2015). Cell intrinsic expression of TLR9 in autoreactive B cells constrains BCR/TLR7-dependent responses. Journal of immunology (Baltimore, Md. : 1950), 194(6), 2504-2512.
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2

Multiparametric Flow Cytometry Analysis

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Cells were kept at 4°C while staining in PBS with 0.5% BSA and 2 mM EDTA in the presence of CD16/32 block (BD clone 2.4G2).
The following antibodies were purchased from BD: CD45.1 BV711 (A20), CD117 BUV395 (2B8), CD135 APC & PE-CF594 (A2F10.1), MHC II V500, BV421, BV510 (M5/114.15.2), CD24 BUV496 (M1/69), CD127 BV421 (SB/199), CD45RA PE (14.8), CD19 BV42 1(1D3). From eBioscience: CD117 PE-Cy7 (2B8), CD317 eFluor450 & APC (eBio927), CD115 PE (AFS98), APC eFluor780 anti-CD11c (N418), MHC II eFluor450 (M5/114/15/2), CD24 PE-Cy7 (M1/69), CD172a APC & PerCP-eFluor710 (P84), Siglec-H PerCP-eFluor710 (eBio440C), eFluor450 Ter119 (Ter119), CD105 eFluor450 (MJ7/18), Irf8 PerCP-eFluor710 (V3GYWCH), CD45R eFluor450 (RA3-6B2), NK1.1 eFluor450 (PK136), CD3 eFluor450 (17A2), Irf4 PE (3E4). From Tonbo Biosciences: CD45.1 PE-Cy7 (A20), CD115 PE (AFS98), CD11c APC-Cy7 (N418). From BioLegend: CD115 BV711 (ASF98), CD45.1 Alexa Fluor 488 (A20), Flag (DYKDDDDK) APC (L5).
Lineage cell depletion kit was purchased from Miltenyi. Cells were analyzed on FACSCanto II or FACSAria Fusion flow cytometers (BD), and data analyzed with FlowJo software (Tree Star).
Grajales-Reyes G.E., Iwata A., Albring J., Wu X., Tussiwand R., Wumesh K., Kretzer N.M., Briseño C.G., Durai V., Bagadia P., Haldar M., Schönheit J., Rosenbauer F., Murphy T.L, & Murphy K.M. (2015). Batf3 maintains Irf8 autoactivation for commitment of a CD8α+ cDC clonogenic progenitor. Nature immunology, 16(7), 708-717.
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3

Multiparametric Flow Cytometry Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were kept at 4°C while staining in PBS with 0.5% BSA and 2 mM EDTA in the presence of CD16/32 block (BD clone 2.4G2).
The following antibodies were purchased from BD: CD45.1 BV711 (A20), CD117 BUV395 (2B8), CD135 APC & PE-CF594 (A2F10.1), MHC II V500, BV421, BV510 (M5/114.15.2), CD24 BUV496 (M1/69), CD127 BV421 (SB/199), CD45RA PE (14.8), CD19 BV42 1(1D3). From eBioscience: CD117 PE-Cy7 (2B8), CD317 eFluor450 & APC (eBio927), CD115 PE (AFS98), APC eFluor780 anti-CD11c (N418), MHC II eFluor450 (M5/114/15/2), CD24 PE-Cy7 (M1/69), CD172a APC & PerCP-eFluor710 (P84), Siglec-H PerCP-eFluor710 (eBio440C), eFluor450 Ter119 (Ter119), CD105 eFluor450 (MJ7/18), Irf8 PerCP-eFluor710 (V3GYWCH), CD45R eFluor450 (RA3-6B2), NK1.1 eFluor450 (PK136), CD3 eFluor450 (17A2), Irf4 PE (3E4). From Tonbo Biosciences: CD45.1 PE-Cy7 (A20), CD115 PE (AFS98), CD11c APC-Cy7 (N418). From BioLegend: CD115 BV711 (ASF98), CD45.1 Alexa Fluor 488 (A20), Flag (DYKDDDDK) APC (L5).
Lineage cell depletion kit was purchased from Miltenyi. Cells were analyzed on FACSCanto II or FACSAria Fusion flow cytometers (BD), and data analyzed with FlowJo software (Tree Star).
Grajales-Reyes G.E., Iwata A., Albring J., Wu X., Tussiwand R., Wumesh K., Kretzer N.M., Briseño C.G., Durai V., Bagadia P., Haldar M., Schönheit J., Rosenbauer F., Murphy T.L, & Murphy K.M. (2015). Batf3 maintains Irf8 autoactivation for commitment of a CD8α+ cDC clonogenic progenitor. Nature immunology, 16(7), 708-717.
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