Western blot analysis of extracts from various cell lines using PVR/CD155 (D3G7H) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
Western blot analysis of extracts from HeLa cells, untreated (lane 1; -), mock treated (lane 2; -), or peptide-N-glycosidase F (PGNaseF)-treated (lane 3; +), using PVR/CD155 (D3G7H) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
|MW (kDa)||60-80 glycosylated 45-50 nonglycosylated|
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. 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
PVR/CD155 (D3G7H) Rabbit mAb recognizes endogenous levels of total PVR (CD155) protein. Based on the protein sequence, this antibody is expected to recognize the α, β, γ, and δ isoforms of PVR (CD155) protein.Species Reactivity:
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg172 of human PVR (CD155) protein.
Poliovirus receptor (PVR, CD155) is an immunoglobulin-like, transmembrane glycoprotein originally described as a mediator of poliovirus attachment to cells and later identified as important in adherens junction formation. Also known as nectin-like 5 (Necl-5), PVR binds nectin-3 and interacts with integrin αvβ3 and PDGFR to regulate integrin clustering and focal contact formation at the leading edge of migrating cells (1,2). Research studies demonstrate that PVR and nectin-3 regulate contact inhibition during cell motility and proliferation in transformed 3T3 cells (3). Additional research indicates that PVR (CD155, Necl-5) expression may play a role in invasiveness of lung adenocarcinoma (4,5). In the immune system, CD155 plays a role in natural killer (NK) cell-mediated cytotoxicity (6).
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