pi 4-Kinase, beta (AA 411-626) antibody

Catalog No. ABIN968667

Product Details

Target: pi 4-Kinase, beta

Binding Specificity: AA 411-626

Reactivity: Dog, Human, Mouse, Rat

Host: Mouse

Clonality: Monoclonal

Application: BioImaging (BI), Western Blotting (WB)

Cross-Reactivity: Dog (Canine), Mouse (Murine), Rat (Rattus)

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
6. Triton is a trademark of the Dow Chemical Company.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Human PI4Kbeta aa. 411-626

Clone: 7-PI4

Isotype: IgG2a

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Comment:

Related Products: ABIN967389, ABIN968535

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 250 μg/mL

Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store undiluted at -20°C.

References

Balla, Downing, Jaffe, Kim, Zólyomi, Catt: "Isolation and molecular cloning of wortmannin-sensitive bovine type III phosphatidylinositol 4-kinases." in: The Journal of biological chemistry, Vol. 272, Issue 29, pp. 18358-66, (1997) (PubMed).

Meyers, Cantley: "Cloning and characterization of a wortmannin-sensitive human phosphatidylinositol 4-kinase." in: The Journal of biological chemistry, Vol. 272, Issue 7, pp. 4384-90, (1997) (PubMed).

Wong, Meyers ddR, Cantley: "Subcellular locations of phosphatidylinositol 4-kinase isoforms." in: The Journal of biological chemistry, Vol. 272, Issue 20, pp. 13236-41, (1997) (PubMed).

Target Details

Target: pi 4-Kinase, beta

Alternative Name: PI4-Kinase beta

Background: Phosphoinositide turnover is a well established mechanism of intracellular signal transduction. Sequential phosphorylation of phosphatidylinositol (PtdIns) results inPtdIns-4-phosphate (PIP) and PtdIns-4,5-bisphosphate (PIP2). Phospholipase C (PLC)hydrolyzes PIP2 to inositol-1,4,5-trisphosphate (IP3) which stimulates release of intracellular Ca2+. PIP is generated by phosphorylation of PtdIns at the D4 position of the inositol ring. This event is mediated by the PtdIns 4-kinases (PI4-K). These enzymes are divided into two types (II and III) based on their size and sensitivity to certain compounds. Although the PI4-Ks are abundantly distributed throughout the cell, activity is found primarily in association with membranous structures. Members of this family contain a lipid kinase unique domain and a C-terminal catalytic domain. Two mammalian PI4-Ks, PI4-Kalpha and PI4-Kß, have been identified. PI4-Kß is homologous to the yeast PI4-K, PIK1. Based on its size and sensitivity to wortmanin (a PI3-K inhibitor), PI4-Kß is classified as a type III enzyme. Although it is found in the cytosol and in association with the Golgi, the specific function of PI4-Kß is yet to be determined.

Molecular Weight: 110 kDa