For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asn101 of human HHLA2/B7-H7 protein.
HERV-H LTR-associating protein 2 (HHLA2, with alternative names of B7-H5 and B7-H7) is a member of the B7 immunoglobulin superfamily (1). HHLA2 protein is constitutively expressed on the surface of human monocytes and is induced on B cells after stimulation with LPS and IFN-γ (1,2). Through interaction with TMIGD2, which is constitutively expressed on all naïve T cells and the majority of natural killer cells, but not on T regulatory cells or B cells, HHLA2 co-stimulates T cells in the context of TCR-mediated activation, enhancing T cell proliferation and cytokine production via an AKT-dependent signaling cascade (2). Contrary to this, HHLA2 has also been shown to inhibit T cell proliferation and cytokine production, suggesting a secondary receptor for HHLA2 that is expressed on activated T cells with co-inhibitory functions (3). Moreover, HHLA2 has been shown to be highly expressed in various types of cancer, and is associated with a poor prognosis (4-10). Further understanding the immunologic functions of the HHLA2 pathway will guide the selection of agents used for cancer immunotherapy, autoimmune disorders, infection, and transplant rejection.
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