CAMP-Dependent Protein Kinase R1 (PKA-R1) (AA 225-381) antibody

Catalog No. ABIN967790

Product Details

Target: CAMP-Dependent Protein Kinase R1 (PKA-R1)

Binding Specificity: AA 225-381

Reactivity: Chicken, Dog, Frog, Human, Mouse, Rat

Host: Mouse

Clonality: Monoclonal

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

Cross-Reactivity: Human, Chicken, Dog (Canine), Frog, 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: Mouse PKA [RI] subunit aa. 225-381

Clone: 18-PKA [RI]

Isotype: IgG2b

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, ABIN968536

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

Chen, Yu, Lee, Chiang, Chao, Huang, Chiong, Huang, Lai, Yang-Yen, Yen: "CREB is one component of the binding complex of the Ces-2/E2A-HLF binding element and is an integral part of the interleukin-3 survival signal." in: Molecular and cellular biology, Vol. 21, Issue 14, pp. 4636-46, (2001) (PubMed).

Orellana, Marfella-Scivittaro: "Distinctive cyclic AMP-dependent protein kinase subunit localization is associated with cyst formation and loss of tubulogenic capacity in Madin-Darby canine kidney cell clones." in: The Journal of biological chemistry, Vol. 275, Issue 28, pp. 21233-40, (2000) (PubMed).

Dohrman, Diamond, Gordon: "Ethanol causes translocation of cAMP-dependent protein kinase catalytic subunit to the nucleus." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 93, Issue 19, pp. 10217-21, (1996) (PubMed).

Cho-Chung: "Role of cyclic AMP receptor proteins in growth, differentiation, and suppression of malignancy: new approaches to therapy." in: Cancer research, Vol. 50, Issue 22, pp. 7093-100, (1990) (PubMed).

Taylor, Buechler, Yonemoto: "cAMP-dependent protein kinase: framework for a diverse family of regulatory enzymes." in: Annual review of biochemistry, Vol. 59, pp. 971-1005, (1990) (PubMed).

Target Details

Target: CAMP-Dependent Protein Kinase R1 (PKA-R1)

Alternative Name: PKA RI

Background: CAMP-dependent Protein Kinase (PKA) is composed of two distinct subunits: catalytic (C) and regulatory (R). Four regulatory subunits have been identified: RIalpha, RIß, RIIalpha, and RIIß. These subunits define type I and II cAMP-dependent protein kinases. Following binding of cAMP, the regulatory subunits dissociate from the catalytic subunits, rendering the enzyme active. Type I and type II holoenzymes have three potential C subunits (Calpha, Cß, or Cgamma). Type II PKA can be distinguished by autophosphorylation of the R-subunits, while type I PKA binds Mg/ATP with high affinity. Most cells express both type I and type II PKAs. Although the Ralpha isoforms are ubiquitously expressed, the Rß isoforms are predominant in nervous and adipose tissues. The levels of expression of the different subunits vary according to cell and tissue type.

Molecular Weight: 48 kDa