Anti cd4 alexa fluor 647 gk1.5

Manufactured by BioLegend

Anti-CD4 Alexa Fluor 647 (GK1.5) is a fluorescently labeled antibody that binds to the CD4 antigen expressed on the surface of helper T cells. The Alexa Fluor 647 dye is used to label the antibody, allowing for detection and analysis of CD4-positive cells by flow cytometry.

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Lab products found in correlation

Propidium iodide, by BioLegend (1 mentions) Anti tcrβ pe cy7, by BioLegend (1 mentions) Novaseq 6000, by Illumina (1 mentions) Facsaria, by BD (1 mentions) Las x software, by Leica (1 mentions) Anti f4 80 alexa fluor 488 bm8, by BioLegend (1 mentions) Tcs sp5 2, by Leica (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Anti gl7 pe, by BioLegend (1 mentions) Fluoromount g with dapi, by Thermo Fisher Scientific (1 mentions) Ihc icc blocking high protein buffer, by Thermo Fisher Scientific (1 mentions) Sp8 inverted 5 channel confocal microscope, by Leica (1 mentions)

Close spelling variants of this product

CD4 Alexa Fluor 647 CD4-Alexa-647 Anti-CD4 (GK1.5, Alexa Fluor 647 CD4 (GK1.5, Alexa 647 Anti-CD4

3 protocols using anti cd4 alexa fluor 647 gk1.5

Single-cell RNA-seq of Regulatory T Cells

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Splenocytes from Foxp3^YFP-creIcos^+/+ (ICOS WT) and Foxp3^YFP-creIcos^fl/fl (ICOS FC) male mice were isolated 6 dpi with NP-OVA/alum and stained with anti-CD4 Alexa Fluor 647 (GK1.5; BioLegend), anti-TCRβ PE-Cy7 (H57-597; BioLegend), and propidium iodide (Thermo Fisher Scientific). Live (PI⁻), conventional (YFP⁻), and regulatory (YFP⁺) CD4⁺TCRβ⁺ T cells were sorted with a BD FACSAria (BD Biosciences) to >95% purity. Sorted conventional and regulatory T cells were mixed in a 1:10 ratio to provide an internal control. A total of 13,500 cells from ICOS WT and ICOS FC mice were sent for library preparation. Libraries were generated using the following components from 10×Genomics: Chromium Next GEM Chip G Single Cell Kit, Chromium Next GEM Single Cell 3′ GEM, Library & Gel Bead Kit v3.1, and Chromium i7 Multiplex Kit. Sequencing was performed by Genome Québec using a NovaSeq 6000 (Illumina) with a flow cell S1 PE28*91.

Panneton V., Mindt B.C., Bouklouch Y., Bouchard A., Mohammaei S., Chang J., Diamantopoulos N., Witalis M., Li J., Stancescu A., Bradley J.E., Randall T.D., Fritz J.H, & Suh W.K. (2023). ICOS costimulation is indispensable for the differentiation of T follicular regulatory cells. Life Science Alliance, 6(4), e202201615.

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Multicolor Immunofluorescence Imaging of Colon Tissue

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Samples of the distal part of the colon were frozen and embedded in Optimal Cutting Temperature compound (Sakura). Frozen tissue sections were fixed in 2% paraformaldehyde for 20 min. To prevent unspecific binding, sections were blocked for 1 h with blocking buffer consisting of 10% fetal calf serum (FCS; Pan-Biotech) and 1% bovine serum albumin (Sigma-Aldrich). Sections were double-stained with anti-F4/80-Alexa Fluor 488 (BM8; BioLegend) and anti-CD4-Alexa Fluor 647 (GK1.5; BioLegend) or rabbit anti-MPO (polyclonal; Abcam) in blocking buffer overnight at 4°C. For MPO staining, sections were additionally incubated with donkey anti-rabbit-IgG-Alexa Fluor 647 (Poly4064; BioLegend) for 2 h at room temperature in blocking buffer. Nuclei were counterstained with Hoechst 33342 (Life Technologies). Images were recorded using the confocal microscope Leica TCS SP5II with Leica LasX software.

Buchele V., Konein P., Vogler T., Kunert T., Enderle K., Khan H., Büttner-Herold M., Lehmann C.H., Amon L., Wirtz S., Dudziak D., Neurath M.F., Neufert C, & Hildner K. (2021). Th17 Cell-Mediated Colitis Is Positively Regulated by Interferon Regulatory Factor 4 in a T Cell-Extrinsic Manner. Frontiers in Immunology, 11, 590893.

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Immunohistochemical Analysis of Mouse Tissues

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H & E staining of sections of spleen, salivary glands, kidneys, lung, small intestine and liver from Eos^fl/fl and Eos cKO mice were performed by HistoServ Labs Inc. (Gaithersburg, MD). Confocal images were taken in the Biological Imaging Section, NIAID, NIH. Histology scores were determined by an independent scorer.
Affixed cryostat sections were dried at 25 °C, fixed in ice-cold acetone for 10 min and dried again at 25 °C. Slides were rehydrated with 1x Tris-buffered saline (TBS) pH 7.6 and placed in a humidifier chamber. Sections were blocked for 2 h at 25 °C with IHC/ICC Blocking High Protein Buffer (eBioscience). Sections were stained overnight at 4 °C in blocking buffer with anti-IgD PerCP e-Fluor 710 (11–26c; eBioscience), anti-CD4 Alexa Fluor 647 (GK1.5; Biolegend), anti-GL7 PE (Biolegend). Slides were then washed in 1x TBS for 5 min with gentle agitation. Slides were mounted with Fluoromount-G with DAPI (eBioscience). Images were visualized and collected using a Leica SP8 inverted 5-channel confocal microscope (Leica) and analyzed in Imaris 8.1 (Bitplane, Oxford Instruments). Images were taken in the Biological Imaging core, NIAID, NIH.

Garofalo S., Metsäranta M., Ellard J., Smith C., Horton W., Vuorio E, & de Crombrugghe B. (1993). Assembly of cartilage collagen fibrils is disrupted by overexpression of normal type II collagen in transgenic mice.. Proceedings of the National Academy of Sciences of the United States of America, 90(9), 3825-3829.

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