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Biotinylated anti rat igg

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Biotinylated anti-rat IgG is a laboratory reagent used for the detection and quantification of rat immunoglobulin G (IgG) in various research and diagnostic applications. It is a secondary antibody conjugated with the biotin molecule, which can be used in conjunction with streptavidin-based detection systems.

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Lab products found in correlation

Efluor 780, by Thermo Fisher Scientific (3 mentions) Lsrfortessa, by BD (3 mentions) Fixation permeabilization kit, by Thermo Fisher Scientific (2 mentions) Fcs express, by De Novo Software (2 mentions) Jes6 5h4, by BD (1 mentions) Pd 1 rmp1 30, by BD (1 mentions) Ebio17b7, by BD (1 mentions) Cell strainer, by BD (1 mentions) Fjk 16, by Thermo Fisher Scientific (1 mentions) Bcl 6, by BD (1 mentions) Live dead fixable near ir dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Bcl 6, by Thermo Fisher Scientific (1 mentions) Facsfortesa, by BD (1 mentions) Foxp3, by Thermo Fisher Scientific (1 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions) Facscanto 2, by BD (1 mentions) Vector novared substrate kit, by Vector Laboratories (1 mentions) Anti cd4, by Agilent Technologies (1 mentions) Anti cd40 1c10, by Thermo Fisher Scientific (1 mentions) Foxp3 stating buffer, by Thermo Fisher Scientific (1 mentions)

7 protocols using biotinylated anti rat igg

1

Multiparameter Flow Cytometry of Immune Cells

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Single-cell suspensions were prepared using standard procedures from spleen, thymus, and mesenteric lymph node (mLN). After red blood cell lysis, cells were blocked with anti-CD16/32 Ab (2.4G2), and stained in FACS staining buffer (2.5% FBS, 0.05% sodium azide in PBS). Fluorochrome-conjugated Abs used were to B220 (RA3-6B2), BCL6 (K112-91), CD4 (RAM4-5), CD8 (53-6.7), CD25 (PC61.5), CD44 (IM7), CD45.1 (A20), CD45.2 (104), CD62L (MEL-14), CD69 (H1.2F3), CD95 (Jo2), CD90.1 (HIS51), CD90.2 (53-2.1), Foxp3 (FJK-16s), Ly-77 (GL7), Neuropilin-1 (761705), PD-1 (RMP1-30), IFN-γ (XMG1.2), IL-2 (JES6-5H4), IL-17a (eBio17B7), and IL-21 (FFA21) purchased from BD Biosciences, eBioscience, BioLegend, and R&D Systems. Follicular T cells were stained as previously described (55 (link)) in a three-step process using purified CXCR5 (2G8) followed by biotinylated anti-rat IgG (Jackson ImmunoResearch), then PerCP5.5-labeled streptavidin in FACS staining buffer on ice. Dead cells were excluded with fixable viability dye (eFluor780; eBioscience). Data were collected on LSRFortessa (BD Biosciences) and analyzed with FlowJo software (Tree Star).
Choi S.C., Hutchinson T.E., Titov A.A., Seay H.R., Li S., Brusko T.M., Croker B.P., Salek-Ardakani S, & Morel L. (2016). The lupus susceptibility gene Pbx1 regulates the balance between follicular helper T cell and regulatory T cell differentiation. Journal of immunology (Baltimore, Md. : 1950), 197(2), 458-469.
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2

Flow Cytometry Analysis of Lymphocytes

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Lymphocytes of the vaccine-dLNs and spleens were harvested by mashing the tissues through cell strainer (BD Falcon). Antibodies for flow cytometry analysis in the study include CD4 (Biolegend, clone RM4-5), CD45.1 (Biolegend, clone A20), PD-1 (Biolegend, clone 29F.1A12), CD44 (eBioscience, clone IM7), B220 (Biolegend, clone), CD19 (Biolegend, clone 1D3/CD19), FAS (BD Biosciences, clone Jo2), PNA (Vector Labs, clone FL-1071), Bcl-6 (BD Biosciences, clone K112-91) and FoxP3 (eBioscience, clone FJK-16s). Stainings of cell surface markers were performed in PBS containing 2% FBS. Staining for CXCR5 was described previously[16] . Briefly, cells were successively stained with purified anti-CXCR5 (BD Biosciences) for 1 hour at 4°C, biotinylated anti-rat IgG (Jackson Immunoresearch) for 30 min on ice and fluorescent-dye conjugated streptavidin (Thermo Fisher) for 30 min on ice. Stainings for FoxP3 and Bcl-6 were performed with the FoxP3/Transcription Factor Staining Buffer Set (eBioscience, 00-5523). Dead cells were stained using LIVE/DEAD Fixable Near-IR Dead Cell Stain Kit (Life Technologies). Flow cytometry data were acquired in a FACSCanto II (BD Biosciences) or a FACSFortesa (BD Biosciences) and analyzed using FlowJo.
Chen X., Lin Y., Yue S., Yang Y., Yang X., He J., Gao L., Li Z., Hu L., Tang J., Wang Y., Tian Q., Hao Y., Xu L., Huang Q., Cao Y, & Ye L. (2023). PD-1/PD-L1 blockade restores tumor-induced COVID-19 vaccine bluntness. Vaccine.
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3

Comprehensive Immune Cell Phenotyping

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Single-cell suspensions were prepared from spleens using standard procedures. After red blood cell lysis, cells were blocked with anti-CD16/32 Ab (2.4G2), and stained in FACS staining buffer (2.5% FBS, 0.05% sodium azide in PBS). Fluorochrome-conjugated Abs used were to detect BCL6 (K112-91), BTK (M4G3LN), CD4 (RAM4-5), CD11c (HL3), CD19 (1D3), CD44 (IM7), CD45 (30-F11), CD69 (H1.2F3), CD80 (16-10A1), CD86 (GL1), Foxp3 (FJK-16s), I-Ab (AF6-120.1), IFN-γ (XMG1.2), IL-6 (MP5-20F3), IL-17A (TC11-18H10.1), IL-21 (FFA21), PD-1 (RMP1-30), PGSL-1 (CD162, clone 2PH1). These antibodies were purchased from BD Biosciences (San Jose, CA, USA), eBioscience (San Diego, CA, USA), or BioLegend (San Diego, CA, USA). CXCR5 was stained in a 3-step process first with the purified antibody (2G8) followed by biotinylated anti-rat IgG (Jackson ImmunoResearch Lab, West Grove, PA, USA) then PerCP-labeled streptavidin. Dead cells were excluded with fixable viability dye (eFluor780, eBioscience). Data were collected on LSRFortessa (BD Biosciences) and analyzed using FCS Express (DeNovo, Glendale, CA, USA) and FlowJo (LLC, Ashland, OR, USA). To analyze intracellular cytokine production, cells were treated with the leukocyte activation cocktail (LKA, BD Biosciences) for 5 h and fixed with the Fixation/Permeabilization kit (eBiosciences).
Choi S.C., Xu Z., Li W., Yang H., Roopenian D.C., Morse HC I.I.I, & Morel L. (2018). Relative Contributions of B Cells and Dendritic Cells from Lupus-Prone Mice to CD4+ T Cell Polarization. Journal of immunology (Baltimore, Md. : 1950), 200(9), 3087-3099.
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4

Immunohistochemical Characterization of Thyroid Tissue

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Frozen or formaldehyde fixed paraffin sections of thyroids were blocked
with 5% BSA in PBS, and endogenous peroxidase was inhibited by
incubation with 0.3% H2O2 for 30 min. Anti-TTF-1 (H-190, Santa Cruz
Biotechnology), anti-CD40 (1C10; eBioscience for frozen sections or C-20 (sc975;
Santa Cruz for paraffin sections), anti-human CD3 (rabbit polyclonal, Dako),
anti-CD4 (clone GK1.5, supernatant) or anti-CD8 (clone 53.6, supernatant) were
used as primary Ab. For staining of frozen sections (CD4, CD8 and CD40),
biotinylated anti-rat IgG (Jackson ImmunoResearch Laboratories Inc., West Grove,
PA) was used as secondary Ab (1:500), followed by avidin-HRP binding using a
Vectastain Elite PK-6100 kit (Vector Laboratories, Burlingame, CA). Peroxidase
activity was visualized using a Vector Nova-Red Substrate Kit (Vector). TTF1,
CD3 and CD40 staining of paraffin sections was done by IDEXX RADIL, Columbia,
MO. They were developed with biotinylated anti-rabbit or anti-goat IgG at
previously determined optimal concentrations followed by avidin-HRP and
visualized using diaminobenzidine tetrahydrochloride (DAB) as the chromogen.
Kayes T.D., Weisman G.A., Camden J.M., Woods L.T., Bredehoeft C., Downey E.F., Cole J, & Braley-Mullen H. (2016). A new murine model of early onset autoimmune thyroid disease/hypothyroidism and autoimmune exocrinopathy of the salivary gland. Journal of immunology (Baltimore, Md. : 1950), 197(6), 2119-2130.
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5

Comprehensive Immune Cell Profiling

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Single-cell suspensions were prepared from spleens and brachial lymph nodes using standard procedures. After red blood cell lysis, cells were blocked with anti-CD16/32 Ab (2.4G2), and stained in FACS staining buffer (2.5% FBS, 0.05% sodium azide in PBS). Fluorochrome-conjugated Abs used were to B220 (RA3-6B2), BCL6 (K112-91), CD3 (145-2C11), CD4 (RAM4-5), CD8 (53-6.7), CD11b (M1/70), CD11c (HL3), CD25 (PC61.5), CD44 (IM7), CD62L (MEL-14), CD69 (H1.2F3), CD95 (Jo2), CD98 (RL388), CD138 (281-2), CD162 (PSLG-1, 2PH1), CD279 (PD-1, RMP1-30), Foxp3 (FJK-16s), I-A/I-E (MHC-II, M5/114.15.2), IL-17A (TC11-18h10.1), Ly6C (HK1.4) and Ly6G (1A8), Ly-77 (GL7), pAKT-s473(SDRNR), pE4-BP (236B4), pS6 (D57.2.2E), and PDCA-1 (129C1), purchased from BD Biosciences, Thermo Fisher, BioLegend, or Cell Signaling Technology. Monocyte/macrophages were gated as CD3– CD11b+Ly6Chi while neutrophils were CD3– CD11bhi Ly6G+. Follicular helper T cells were stained as previously described (3 (link)) in a three-step process using purified CXCR5 (2G8) followed by biotinylated anti-rat IgG (Jackson ImmunoResearch Laboratory) then PerCP-labeled streptavidin in FACS staining buffer on ice. For the intracellular staining, cells were fixed and permeabilized with FOXP3 stating buffer (Thermo Fisher) according to the manufacturer's protocol.
Abboud G., Choi S.C., Kanda N., Zeumer-Spataro L., Roopenian D.C, & Morel L. (2018). Inhibition of Glycolysis Reduces Disease Severity in an Autoimmune Model of Rheumatoid Arthritis. Frontiers in Immunology, 9, 1973.
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6

Multiparameter Flow Cytometry of Murine Immune Cells

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Spleen cells were stained with anti-Fc II/III receptor mAb 2.4G2 for 15 min at 4°C. Surface staining was performed on splenic single cell suspension with fluorescently-labeled or biotinylated antibodies against CD4 (BD Pharmingen 1:200), CD8 (BD Pharmingen 1:200), B220 (BD Pharmingen 1:200), CXCR5 (BD Pharmingen 1:1000), PD-1 (eBioscience 1:200), FAS (BD Pharmingen 1:500), IgD (Southern Biotechnology Associates 1:1000), CD138 (Biolegend 1:1000), CD45.1 (BD Pharmingen 1:400), CD45.2 (BD Pharmingen 1:400), OX40 (BD Pharmingen 1:200) and fluorescently-labeled PNA (Vector laboratories 1:1000). Streptavidin- APC and -PE were used to stain biotinylated antibodies. CXCR5 staining was done as a three-step stain, as published (2 (link)) using purified rat anti-mouse CXCR5 (BD Pharmingen 1:1000) followed by biotinylated anti-rat IgG (Jackson ImmunoResearch Laboratories 1:1000) in PBS/0.5% BSA/2% FCS/2% normal mouse serum on ice. For tertiary staining of CXCR5, PE or APC-labeled streptavidin (Caltag Laboratories) was used in PBS/0.5% BSA/2% FCS. BCL6 was stained intracellularly after permeabilization with Cytofix/Cytoperm (BD). Samples were acquired on LSR II (BD Biosciences) and analyzed using FlowJo (Tree Star).
Tahiliani V., Hutchinson T.E., Abboud G., Croft M, & Salek-Ardakani S. (2016). OX40 Co-operates with ICOS to Amplify Follicular T Helper Cell Development and Germinal Center Reactions During Infection. Journal of immunology (Baltimore, Md. : 1950), 198(1), 218-228.
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7

Comprehensive T-cell Phenotyping by Flow Cytometry

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Single-cell suspensions were prepared from spleens using standard procedures. After red blood cell lysis, cells were blocked with anti-CD16/32 Ab (2.4G2), and stained in FACS staining buffer (2.5% FBS, 0.05% sodium azide in PBS). Fluorochrome-conjugated Abs used were to B220 (RA3-6B2), BCL6 (K112-91), CD4 (RAM4-5), CD25 (PC61.5), CD44 (IM7), CD62L (MEL-14), CD69 (H1.2F3), CD95 (Jo2), CD122 (TM-b1), CD152 (UC10-4B9), CD154 (MR1), FOXP3 (FJK-16s), ICOS (15F9), IFN-γ (XMG1.2), IL-17A (TC11-18H10.1), Ly-77 (GL7), PD-1 (RMP1-30), purchased from BD Biosciences, eBioscience, and BioLegend. Follicular T cells were stained as previously described (55) in a three-step process using purified CXCR5 (2G8) followed by biotinylated anti-rat IgG (Jackson ImmunoResearch) and PerCP5.5-labeled streptavidin in FACS staining buffer on ice. Dead cells were excluded with fixable viability dye (eFluor780, eBioscience). Data were collected on LSRFortessa (BD Biosciences) and analyzed with FlowJo (Tree Star) or FCS Express (DeNovo) software. IFN-γ and IL-17A production were analyzed in cells treated with the leukocyte activation cocktail (BD Biosciences) for 5 h and the Fixation/Permeabilization kit (eBiosciences).
Yin Y., Choi S.C., Xu Z., Zeumer L., Kanda N., Croker B.P, & Morel L. (2015). Glucose oxidation is critical for CD4+ T cell activation in a mouse model of systemic lupus erythematosus. Journal of immunology (Baltimore, Md. : 1950), 196(1), 80-90.
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