Analysis Note

ControlMouse thymocyte tissue lysate

Application

Western Blot (Snap i.d.) Analysis: 2 µg/mL from a previous lot detected Themis in 10 µg of mouse thymocyte tissue lysate.

Flow Cytometry Analysis: A previous lot was used by an independent laboratory in FC. (Vasily Rybakin, PhD., Gascoigne lab, Scripps Research Institute, La Jolla, CA

Western Blot Analysis: A previous lot of this antibody was used to detect the presence and absence of Themis in Themis KO or WT thymocytes (Fu, G., et al. (2013) Nature. doi: 10.1038/nature12718)

Immunopreciptiation Analysis: A previous lot of this antibody was used to immunoprecipitate Themis in WT thymocytes but not in Themis KO thymocytes (Fu, G., et al. (2013) Nature. doi: 10.1038/nature12718)

Detect Themis using this Anti-Themis Antibody validated for use in WB, FC.

General description

Thymocyte-expressed molecule involved in selection (Themis) is a highly conserved protein that has been shown to serve a key role in regulating T-cell development through TCR signaling, especially through the moderation of phosphorylation and calcium influx. It is also a critical component in the later stages of thymocyte development through the regulation of positive/negative T-cell selection, and is required for integration and maintenance of signals necessary to T-cell lineage commitment and maturation. Themis expression is observed in thymus, spleen, thymocytes between Pre-TCR and checkpoints positive in selectin, and at low levels in mature T-cells.

Immunogen

KLH-conjugated linear peptide corresponding to the C-terminus of mouse Themis.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Quality

Evaluated by Western Blot in mouse thymocyte tissue lysate.

Western Blot Analysis: 0.1 µg/mL of this antibody detected Themis in 10 µg of mouse thymocyte tissue lysate.

Specificity

This antibody recognizes Themis at the C-terminus.

Target description

~72 kDa observed.
UniProtKB/Swiss-Prot Q8BGW0 (THMS1_MOUSE) entry describes 5 isoforms produced by alternative splicing: Isoform 1 at 72.780 kDa, Isoform 2 at 69.544 kDa, Isoform 3 at 68.293 kDa, Isoform 4 at 59.082 kDa, and Isoform 5 at 30.040 kDa