|H M R Mk B Pg||Endogenous||50||Rabbit IgG|
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, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
PIP4K2A (D83C1) Rabbit mAb recognizes endogenous levels of total PI 5-P 4-kinase type-2 alpha (PIP4K2A) protein. This antibody does not cross-react with PIP4K2B or PIP4K2C and is not predicted to cross-react with type I PIP5Ks or PIKfyve.
Human, Mouse, Rat, Monkey, Bovine, Pig
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human PIP4K2A protein.
Phosphatidylinositol 5-phosphate 4-kinase type-2 alpha (PtdIns 4-Kinase type II alpha, PIP4K2A), is one of three known members of the type II PIP kinase family, consisting of PIP4K2A, PIP4K2B, and PIP4K2C. Each catalyzes the phosphorylation of phosphatidylinositol 5-monophosphate (PI 5-P) to form phosphatidylinositol 4,5-bisphosphate (PI 4,5-P2). Originally thought to be a PI 4-P 5-Kinase (1,2), PIP4K2A was subsequently shown to phosphorylate the 4-position of PI 5-P, thus defining a new family of lipid kinases (3). Ubiquitously expressed with highest levels in the brain, mutations in PIP4K2A have been described in patients with Schizophrenia and other neuronal disorders (4-8).
The levels of PI 5-P change significantly in response to physiological and pathological stimuli (5-12), as well as cell transformation with nucleophosmin anaplastic lymphoma tyrosine kinase (13). In contrast, hypoosmotic shock and histamine decrease cellular levels of PI 5-P (14,15). PIP4K2A has been hypothesized to play a role in suppressing mitogen-dependent increases in PI 5-P in response to DNA damage and cellular stress (16-18). PIP4K2A regulates the levels of PI 5-P in the nucleus by converting the PI 5-P to PI 4,5-P2, thus preventing PI 5-P from interacting with and regulating the ability of ING2 to activate p53 and p53-dependent apoptotic pathways (19). PIP4K2A has been shown to form a heterodimer with PIP4K2B resulting in its recruitment to the nucleus. Interestingly, PIP4K2A is 2000-fold more active than PIP4K2B in this context, suggesting that the two lipid kinases act in tandem, with PIP4K2B acting as the targeting subunit and PIP4K2A the catalytic component (18). PIP4Ks may also play a role in lipid vesicle formation and/or Golgi homeostasis (20).
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|5527S||100 µl (10 western blots)||$ 255.0|