Spleens were frozen in OCT at −80°C before sectioning in a Microm‐HM‐525 cryostat. Tissue sections were fixed in acetone for 5 min and blocked with 3% BSA/PBS. Sections were stained overnight with conjugated antibodies: FITC‐conjugated anti‐B220 or anti‐GL7, and biotinylated anti‐CD4 (all Biolegend) with a secondary stain of AlexaFluor‐594‐conjugated streptavidin (Thermo Life Technologies, MA, USA). Sections were mounted with Fluoromount (Southern Biotech, AL, USA) and imaged with a fluorescence BZ‐X850 microscope with a BZ‐X software (Keyence, Osaka, Japan).
Anti gl7
Manufactured by BioLegend
Sourced in United States
Anti-GL7 is a laboratory reagent used to detect the GL7 antigen. GL7 is a glycolipid expressed on activated germinal center B cells and is involved in the immune response. Anti-GL7 can be used in flow cytometry and other immunological assays to identify and analyze GL7-expressing cells.
Automatically generated - may contain errors
Lab products found in correlation
Anti b220, by BioLegend (4 mentions)
Anti cd86, by BioLegend (3 mentions)
Anti cd11c, by BioLegend (3 mentions)
Anti pd 1, by BioLegend (2 mentions)
Anti cd40, by BioLegend (2 mentions)
Anti f4 80, by BioLegend (2 mentions)
Anti cd95, by BioLegend (2 mentions)
Anti cxcr5, by BioLegend (2 mentions)
Anti cd4, by BioLegend (2 mentions)
Anti igd, by BioLegend (2 mentions)
Anti cd19, by BioLegend (2 mentions)
Fitc conjugated anti b220, by BioLegend (1 mentions)
Bz x software, by Keyence (1 mentions)
Fluoromount, by Southern Biotech (1 mentions)
Anti cd80, by BioLegend (1 mentions)
Anti cd38, by BioLegend (1 mentions)
Anti cd69, by BioLegend (1 mentions)
Anti mhcii, by BioLegend (1 mentions)
Anti gr1, by BioLegend (1 mentions)
Anti cd3, by BioLegend (1 mentions)
7 protocols using anti gl7
1
Immunofluorescent Tissue Staining Protocol
2
Comprehensive Antibody Validation for Immunoblotting and Flow Cytometry
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Anti-mouse β-actin (AF0003; 1:1,000 dilution), Horseradish peroxidase (HRP)-anti-rabbit immunoglobulin G (IgG) (A0208; 1:3,000 dilution) and HRP-anti-mouse IgG (A0216; 1:3,000 dilution) were purchased from Beyotime Institute of Biotechnology (Haimen, China). Anti-ACK1 (PA5-102625) antibody was purchased from Invitrogen (Carlsbad, CA, USA). Anti-p-p65 (9246), anti-p65 (9242), anti-p-Erk (9101), anti-Erk (9107), anti-p-JNK (9251), anti-JNK (9251), anti-p-p38 (4511) and anti-p38 (9228) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). For flow cytometry purposes, fluorochrome-conjugated anti-B220 (Cat#: 103206), anti-CD4 (Cat#: 100406), anti-CD40 (Cat#: 124610), anti-F4/80 (Cat#: 123108), anti-CD86 (Cat#: 105012), anti-CD11c (Cat#: 117310), anti-GL7 (Cat#: 144608), anti-CD95 (Cat#: 152604), anti-CXCR5 (Cat#:145506), and anti-PD-1 (Cat#:135206) antibodies were purchased from BioLegend. Unless specifically listed otherwise, all flow cytometry antibodies were stained at a 1:100 dilution.
3
Comprehensive Immune Cell Profiling
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The cells were resuspended in ice-cold PBS containing 1% FBS and were stained with anti-CD3, anti-CD4, anti-CD8, anti-B220, anti-CD38, anti-CD138, anti-PD-1, anti-CXCR5, anti-CD62L, anti-OX40, anti-IgD, anti-CD95, anti-GL7, anti-CD11c, anti-CD80, anti-CD86, anti-MHC II, anti-CD40, anti-CD69, anti-CD11b, anti-Gr-1, and anti-F4/80 antibodies (Biolegend). Intracellular staining of IFN-γ, TNF-α, IL-17a, Helios, and Foxp3 was performed after 4 h of stimulation with PMA and ionomycin following a standard protocol (BD Biosciences). The results were obtained on a BD FACS Fortessa flow cytometer and were analyzed using FlowJo software.
4
Multiparametric Immune Cell Profiling
5
Multicolor Immunofluorescence of Lung Tissue
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Lungs frozen in Tissue-Tek O.C.T. compound (Sakura Finetek) were sectioned using a CM1850 cryostat (Leica). 30-µm frozen sections were blocked for 30 min with Background Buster (Innovex Biosciences) at room temperature, stained 6 h at 4°C in a humidified chamber with anti-CD8α (BD), anti-GL-7 (BioLegend), and anti-B220 (BioLegend) in PBS with 2% goat serum. Alternatively (for lymphoid structures in the lung), whole mount imaging was performed on 250 µm scalpel cut lung sections that were then fixed with 2% PFA (1 h at 4°C), blocked, and stained with the antibody cocktail mentioned above and anti-CD4 Ab (BD). Immunofluorescence confocal microscopy was performed with the Zeiss 780 laser scanning microscope (Carl Zeiss; air objective 20× Plan-Apochromat with NA [numerical aperture] 0.5 or water objective 10x Plan-Apochromat with NA 0.30) using multichannel frame scans. Image processing was performed using Imaris 8.1 software. Images were analyzed using LSM5 image browser and cells in three to four images from each mouse were counted using ImageJ (National Institutes of Health) cell counter plugin.
6
Comprehensive B Cell Immunophenotyping
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Bone marrow, spleen, mesenteric lymph node (mLN) and Peyer’s Patch (PP) cells were isolated as described previously (26 (link)). In brief, cells were stained with anti-B220 (BioLegend), anti-CD11b (BD Biosciences), anti-CD43 (eBioscience), anti-CD24 (BioLegend), anti-BP1 (eBioscience), anti-IgD (BioLegend), anti-IgM (BioLegend), anti-CD93 (eBioscience) and anti-CD23 (BioLegend) for B cell development staining. For B1 and B2 cell development staining, cells were stained with anti-B220 (BioLegend), anti-CD19 (BioLegend), anti-CD43 (eBioscience), anti-CD23 (BioLegend) and anti-CD5 (BioLegend). For FOB and MZB cell staining, cells were stained with anti-B220 (BioLegend), anti-CD19 (BioLegend), anti-CD21 (BD Biosciences) and anti-CD23 (BioLegend).Cells were stained with anti-B220 (BioLegend), anti-Gl7 (BioLegend), anti-CD95 (Fas) (BD Biosciences), anti-CD86 (BioLegend), and anti-CXCR4(BioLegend) for GC B cell staining and cells were stained with anti-B220 (BioLegend), anti-MHC II (eBioscience) and anti-CD86 (BioLegend) for B cell activation staining.
7
Comprehensive Immune Cell Profiling
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Cells were labeled with anti-CD3 (APC-C7, 17A2 clone), anti-CD27 (APC, LG.3A10 clone), anti-FOXP3 (PE, MF23 clone), anti-B220 (PerCP, RA3-6B2 clone), anti-CD8a (APC-H7 536.7 clone), anti-IL-21R (PE, 4A9 clone) and anti-CD19 (APC-Cy7, ID3 clone) from BD Biosciences; anti-CD4 (eFluor710-PerCP, clone RM4-5), anti-PD-1 (PE-Cyanine7, clone J43), anti-Bcl-6 (PE, mgl191E clone), anti-CXCR5 (APC, clone SPRCL5), anti-ICOS (FITC, clone 7E17G9), anti-CD45 (FITC, clone 30-F11) and anti-CD274 (PE, clone MIH5) from eBioscience; anti-CD25 (PE, PC61 clone), anti-GL7 (FITC, GL7 clone), anti-CD11c (PE/Cy7 N418 clone) and anti-Ki67 (PE or PerCP, clone 16A8) from BioLegend. Viability was determined with Fixable Viability Dye eFluor 780 or 450 from eBioscience. For intracellular staining, we used Foxp3 staining buffer set (eBioscience). Cells were analyzed using an FACSCanto II (BD Biosciences) with the FACSDiva software (BD Biosciences). Sorting was performed in an FASCAria (BD Biosciences).
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