αcd3 145 2c11

Manufactured by BioLegend

αCD3 (145-2C11) is a hamster monoclonal antibody that recognizes the CD3 complex on T cells. CD3 is a T cell co-receptor that is essential for T cell activation and signaling.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

αCD3 mAb (145–2c11, (2 citations)

3 protocols using αcd3 145 2c11

T Cell Activation and Lineage Polarization

Check if the same lab product or an alternative is used in the 5 most similar protocols

Purified T cells were stimulated with plate-bound 5 µg/ml αCD3 (145-2C11; BioLegend) and 5 µg/ml αCD28 (37.51; Tonbo), polarized to the indicated lineages, and harvested at the designated time points. Cells were washed twice with PBS and lysed with RIPA buffer. Protein concentrations were quantified using the Bradford Assay, and equal amounts of protein were resolved on 4–12% Bis-Tris Plus gels (Thermo Fisher Scientific), transferred to PVDF membrane (EMD Millipore), and blotted for BCAP (1 µg/ml; 501813; R&D Systems) or H3 histone (1:3,000; Cell Signaling). Blots were then probed with appropriate secondary antibodies conjugated to fluorophores (IRDye 800CW goat anti–mouse, IRDye 680RD goat anti–rabbit), and membranes were imaged using LI-COR Odyssey CLx.

Deason K., Troutman T.D., Jain A., Challa D.K., Mandraju R., Brewer T., Ward E.S, & Pasare C. (2018). BCAP links IL-1R to the PI3K–mTOR pathway and regulates pathogenic Th17 cell differentiation. The Journal of Experimental Medicine, 215(9), 2413-2428.

+ Open protocol

+ Expand

T Cell Lineage Specification Protocols

Check if the same lab product or an alternative is used in the 5 most similar protocols

For polarizations using plate-bound αCD3 + αCD28, tissue culture plates were coated with 5 µg/ml αCD3 (145-2C11; BioLegend) and 5 µg/ml αCD28 (37.51; Tonbo), incubated at 37°C for 4 h, and washed three times immediately before use. For differentiation to Th17 cells, naive CD4⁺CD62L^highCD44^low T cells were incubated with IL-6 (20 ng/ml; Peprotech), hTGF-β (5 ng/ml; Peprotech), α IL-4 (10 µg/ml; 11B11; BioLegend), and αIFN-γ (10 µg/ml; XMG1.2; BioLegend), with or without IL-1β (2–10 ng/ml; Peprotech). For differentiation to Th1 cells, naive CD4⁺CD62L^highCD44^low T cells were incubated with IL-12 (10 ng/ml; Peprotech), IL-2 (50 U/ml; eBioscience), and α IL-4 (10 µg/ml; 11B11; BioLegend), with or without IL-18 (0.3–3 ng/ml; Gibco). Th2 cell differentiation was achieved by incubating naive CD4⁺CD62L^highCD44^low T cells with IL-4 (4 ng/ml; Peprotech), IL-2 (50 U/ml; eBioscience), and αIFN-γ (10 µg/ml; XMG1.2; BioLegend).

+ Open protocol

+ Expand

Isolation and Activation of Murine T-cells and Dendritic Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total leukocytes were isolated from LN and spleen from 8 to 12 week old mice as previously described [85 (link)]. CD4+ T-cells were isolated from the pooled cell suspensions using an EasySep™ Mouse CD4+ T Cell Isolation Kit from Stemcell Technologies (Vancouver, Canada) following the manufacturer’s instructions. Foxp3+ Treg induction protocols were as previously described [52 (link)]. Polyclonal T-cell activation was performed as previously described [87 (link)], pre-coating plates with 10 μg/ml αCD3 (145–2C11, Biolegend) in PBS overnight at 4°C. Coated plates were washed and cells were plated with 1 μg/ml soluble αCD28 antibody (37.51, Biolegend). Bone marrow-derived dendritic cells (BMDC) were generated as previously described [86 (link)]. DCs were activated overnight with 1 ug/ml LPS (Sigma) along with OVA peptide 323–339 (Invivogen) for antigen specific activation. For polyclonal activation via APC, BMDC Fc receptors were pre-loaded with 2.5 ug/ml αCD23 (Biolegend) and .5 ug/ml αCD28 (Biolegend) for two hours before culturing with target CD4+ T-cells. Antigen processing assays were performed according to manufacturer’s protocol for DQ-OVA (Thermo Fisher Scientific). A phagocytosis assay using these cells in conjunction with pHrodo-labelled particles was performed as previously described [86 (link)].

Marshall P.L., Nagy N., Kaber G., Barlow G.L., Ramesh A., Xie B.J., Linde M.H., Haddock N.L., Lester C.A., Tran Q.L., de Vries C., Hargil A., Malkovskiy A., Gurevich I., Martinez H.A., Kuipers H.F., Yadava K., Zhang X., Evanko S.P., Gebe J.A., Wang X., Vernon R.B., de la Motte C., Wight T.N., Engleman E.G., Krams S.M., Meyer E, & Bollyky P.L. (2020). Hyaluronan synthesis inhibition impairs antigen presentation and delays transplantation rejection. Matrix biology : journal of the International Society for Matrix Biology, 96, 69-86.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!