|H M R Mk||Endogenous||50||Rabbit|
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
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.
γ-Tubulin Antibody recognizes endogenous levels of total γ-tubulin protein.
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human γ-tubulin protein. Antibodies are purified by protein A and peptide affinity chromatography.
Globular tubulin subunits comprise the microtubule building block, with α/β-tubulin heterodimers forming the tubulin subunit common to all eukaryotic cells. As a critical part of the microtubule-organizing center (MTOC), the third member of the tubulin superfamily, γ-tubulin, is required for microtubule nucleation as well as centrosome duplication and spindle assembly (1,2, reviewed in 3). γ-tubulin forms complexes of two different sizes: γ-tubulin small complex (γ-TuSC) and the larger γ-tubulin ring complex (γ-TuRC). Each complex consists of a number of γ-tubulin complex proteins (GCPs) with γ-tubulin itself being considered GCP1. GCP2-6 all share sequence similarity in 5 different regions and it is thought that these areas could play a role in the proper folding of the proteins (4). γ-TuSC is composed of two γ-tubulin molecules as well as GCP2 and GCP3. γ-TuRC is made up of a ring of multiple copies of γ-TuSC in addition to GCP4, 5, and 6. Another protein, GCP-WD/NEDD1, which lacks sequence similarity with the other GCPs, associates with the γ-TuRC. GCP-WD/NEDD1 has been shown to regulate localization of the γ-TuSC to spindles and centrosomes (5-8). In mammals, phosphorylation of γ-tubulin at Ser131 by SADB controls the activity of the γ-TuRC. The hypothesis is that this phosphorylation stabilizes the protein in a conformation that stimulates centrosome amplification (9).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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|5886S||100 µl (10 western blots)||$ 255.0|