Cd4 clone l3t4

Manufactured by Thermo Fisher Scientific

The CD4 (clone L3T4) is a laboratory reagent used for the identification and enumeration of CD4+ T cells. It is a mouse monoclonal antibody that binds specifically to the CD4 cell surface antigen. This product is intended for research use only and its performance characteristics have not been established for diagnostic or clinical procedures.

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CD4 (L3T4; (2 citations)

2 protocols using cd4 clone l3t4

Isolation and Characterization of Adipose Tissue Stromal Vascular Fraction

Cited 1 time

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Visceral adipose tissue (VAT) was dissociated with collagenase I in KRHB buffer for ~30–45 minutes at 37°C as described [30 (link)]. Digested tissue was filtered through a 100 μm cell strainer. Filtrate was centrifuged at 500 x g for 5 min, the pellet resuspended in KRHB, and the process repeated 4 times. The final pellet was collected as the SVF. Cell labeling for flow cytometry was performed as described [31 (link)]. Cells were analyzed on a FACSCalibur cytometer or an LSRII cytometer (BD Biosciences), and data were analyzed using FlowJo (Treestar). The gating schemes are given in S1 Fig and represent the gates used for quantitation of population percentages and cell numbers (obtained using CountBright Absolute Counting Beads according the manufacturer’s instructions; Life Technologies). The following antibodies were used: CD45.2 (clone 104), CD3 (17A2), CD11b (clone M1/70), MHC-II (clone M5/114.15.2), F4/80 (clone BM8), CD4 (clone L3T4), and CD8 (clone 53–6.7), all from eBioscience, and anti-mouse CD25 (clone PC61) from BioLegend.

Bhagwandin C., Ashbeck E.L., Whalen M., Bandola-Simon J., Roche P.A., Szajman A., Truong S.M., Wertheim B.C., Klimentidis Y.C., Ishido S., Renquist B.J, & Lybarger L. (2018). The E3 ubiquitin ligase MARCH1 regulates glucose-tolerance and lipid storage in a sex-specific manner. PLoS ONE, 13(10), e0204898.

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Comprehensive Immunophenotyping of Influenza Infection

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Lungs were harvested at the indicated time points and RNA was extracted by TRI-Reagent (Molecular Research Center). Viral loads were determined by RT-PCR using influenza virus specific primers as previously described (30 (link)). Cells were stained as previously described (31 (link)) using monoclonal antibodies against CD69 (clone H1.2F3), Thy1.1 (clone HIS51), Thy1.2 (clone 53-2.1), CD4 (clone L3T4), CD45.1 (clone A20), CD45.2 (clone 104), CD62L (clone MEL-14), CD44 (clone 1M7), CD127 (clone A7R34), KLRG1 (clone 2F1), IFN-γ (clone XMG1.2), TNF-α (clone MP6-XT22), and Granzyme-B (clone NGZB) (all from eBioscience, San Diego, CA), CD8α (clone 53-6.7, BD Biosciences, San Jose, CA), CD25 (clone PC61, BD Biosciences), and H-2^b MHC class I tetramers loaded with immunodominant influenza virus nuclear protein (NP) epitope NP_(366-374) or OVA_(257-264) peptide. Anti-CD16/32 (Fc Block, clone 2.4G2) (BD Biosciences) was used in all stains. For intracellular cytokine and granule staining, GolgiPlug (BD Biosciences), Cytofix/Cytoperm buffer (eBioscience) and Perm/Wash buffer (eBioscience) were used according to the manufacturer’s instructions. Samples were analyzed using a FACSAria flow cytometer (BD Biosciences). All data were analyzed using FlowJo software (Treestar).

Gracias D.T., Boesteanu A.C., Fraietta J.A., Hope J.L., Carey A.J., Mueller Y.M., Kawalekar O.U., Fike A.J., June C.H, & Katsikis P.D. (2016). Phosphoinositide 3-Kinases p110δ isoform regulates CD8+ T cell responses during acute viral and intracellular bacterial infections. Journal of immunology (Baltimore, Md. : 1950), 196(3), 1186-1198.

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