Western blot analysis of extracts from various human cells using TRBC1/TCRβ constant region 1 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human TRBC1/TCRβ constant region 1 protein (hTRBC1-Myc/DDK; +) and Myc/DDK-tagged full-length human TRBC2/TCRβ constant region 2 protein (hTRBC2-Myc/DDK; +), using TRBC1/TCRβ constant region 1 Antibody (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
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 datasheet or 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 November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised October 2016
Protocol Id: 10
TRBC1/TCRβ constant region 1 Antibody recognizes endogenous levels of total TRBC1/TCRβ constant region 1 protein. This antibody does not cross-react with TRBC2/TCRβ constant region 2 protein.Species Reactivity:
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human TRBC1/TCRβ constant region 1 protein. Antibodies are purified by peptide affinity chromatography.
The αβ T Cell Receptor (TCR) is a heterodimer composed of an alpha chain and a beta chain expressed on the surface of T cells (1,2). It detects peptide antigen presented by major histocompatibility complex (MHC) molecules (1,2). Detection of cognate antigen leads to activation of the TCR signaling pathway through the CD3 signaling chains that associate with the TCR (1). The TCR chains are made up of a constant region and variable regions that get rearranged during T cell development in the thymus (3). There are two TCRβ constant genes, TRBC1 and TRBC2 (3). A population of normal T cells is polyclonal and contains a mix of TRBC1- and TRBC2-expressing T cells, while T cell malignancies are monoclonal and all express the same TCR which either contains TRBC1 or TRBC2 (4). Therefore, it is possible that T cell malignancies could be identified and targeted with reagents or therapies that can distinguish between TRBC1 and TRBC2 (4).
Explore pathways + proteins related to this product.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Tween is a registered trademark of ICI Americas, Inc.