Kidney Androgen Regulated Protein (KAP) (AA 1-212) antibody

Details for Product No. ABIN967909
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Antigen
Synonyms AW146435, FKSG22
Epitope
AA 1-212
(2), (1)
Reactivity
Human
(2), (2)
Host
Mouse
(2)
Clonality (Clone)
Monoclonal ()
Application
Western Blotting (WB), Immunohistochemistry (IHC), BioImaging (BI)
(2)
Pubmed 2 references available
Catalog no. ABIN967909
Quantity 50 µg
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Immunogen Human KAP
Clone DB7
Isotype IgG1
Cross-Reactivity Chicken, Dog (Canine), Frog, 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 Purified from tissue culture supernatant or ascites by affinity chromatography.
Purity Purified
Alternative Name KAP
Background Proteins that associate with cyclin-dependent kinases may function in cell cycle regulation. Therefore, a two-hybrid screen system was used to isolate cDNA which encode proteins that can interact with Cdk2. One of these was the dual-specificity protein phosphatase known as KAP (Cdk associated phosphatase). KAP contains the HCXXXXGR motif characteristic of protein tyrosine phosphatases. It utilizes substrates that contain either phosphotyrosine or phosphoserine residues. In addition to binding Cdk2, KAP also associates with Cdc2. KAP does not require a cyclin subunit for binding to Cdk2 and Cdc2. The association of KAP with these two Cdks suggests that KAP may play a role in cell cycle control, possibly by regulating the phosphorylation status of a cdk or of a Cdk-associated protein.
Molecular Weight 34 kDa
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, ABIN968619

Restrictions For Research Use only
Format Liquid
Concentration 250 µg/ml
Buffer Aqueous buffered solution containing BSA, glycerol.
Preservative Sodium azide
Storage -20 °C
Supplier Images
anti-Kidney Androgen Regulated Protein (KAP) (AA 1-212) antibody Western blot analysis of KAP on a MDCK lysate (ABIN968619). Lane 1: 1:5000, lane 2: 1:10000, lane 3: 1:20000 dilution of the KAP antibody.
anti-Kidney Androgen Regulated Protein (KAP) (AA 1-212) antibody (2) Immunofluorescent staining of A549 (ATCC CCL-185) cells. Cells were seeded in a 96 well imaging plate at ~ 10 000 cells per well. After overnight incubation, cells were stained using the Triton™ X-100 perm protocol and the anti-KAP antibody. The second step reagent was FITC goat anti mouse Ig. The image was taken on a BD Pathway™ 855 Bioimager using a 20x objective. This antibody also stained HeLa (ATCC CCL-2) and U-2 OS (ATCC HTB-96) cells and can be used with either fix/perm protocol.
anti-Kidney Androgen Regulated Protein (KAP) (AA 1-212) antibody (3) anti-Kidney Androgen Regulated Protein (KAP) (AA 1-212) antibody (Image 3)
Product cited in: Hannon, Casso, Beach: "KAP: a dual specificity phosphatase that interacts with cyclin-dependent kinases." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 91, Issue 5, pp. 1731-5, 1994 (PubMed).

Lee, Reimer, Fang et al.: "Overexpression of kinase-associated phosphatase (KAP) in breast and prostate cancer and inhibition of the transformed phenotype by antisense KAP expression." in: Molecular and cellular biology, Vol. 20, Issue 5, pp. 1723-32, 2000 (PubMed).

Hosts (2)
Reactivities (2), (2)
Applications (2)
Epitopes (2), (1)
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