Western blot analysis of extracts from HeLa, Saos-2, and MCF7 cells using Kindlin-2 Antibody.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human kindlin-1 protein (hKindlin-1-Myc/DDK; +), kindlin-2 protein (hKindlin-2-Myc/DDK; +) or kindlin-3 protein (hKindlin-3-Myc/DDK; +) protein, using Kindlin-2 Antibody (upper) or Myc-Tag (71D10) Rabbit mAb #2278 (lower).
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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
Kindlin-2 Antibody recognizes endogenous levels of total kindlin-2 protein. This antibody does not cross-react with kindlin-1 or kindlin-3 proteins.Species Reactivity:
Human, Mouse, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asn502 of human kindlin-2 protein. Antibodies are purified by protein A and peptide affinity chromatography.
The kindlin family of focal adhesion proteins is involved in multiple biological processes, including integrin signaling, adhesion, migration, angiogenesis, differentiation, and mitotic spindle formation (1,2). Kindlin family members 1, 2, and 3 (FERM1, FERM2, and URP2) are differentially expressed in tissues. Kindlin-1 is primarily expressed in epithelial cells, kindlin-2 is ubiquitously expressed, and kindlin-3 expression is restricted to the hematopoietic system (3).
Kindlin-2 regulates integrin β3 mediated adhesion and migration (4,5) and contributes to Wnt signaling through interaction with β-catenin and activation of Wnt target genes (6).
Research studies indicate that kindlin-2 expression in pancreatic and gastric cancers may be indicative of poor prognosis (7,8) and that kindlin-2 may support invasiveness in breast cancer (9). Additional studies show that expression of kindlin-2 in prostate cancer cells can protect cells from cisplatin-induced cell death (10).
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