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FITC Mouse Anti-PKA [RI] 18/PKA [RI] RUO

FITC Mouse Anti-PKA [RI] 

Clone  18/PKA [RI]   (RUO)

  • Brand BD Transduction Laboratories™
  • Concentration 250 µg/ml
  • Isotype Mouse IgG2b
  • Reactivity Human (QC Testing)  Mouse, Rat, Dog, Chicken, Frog (Tested in Development) 
  • Application Bioimaging (Routinely Tested) 
    Immunofluorescence (Tested During Development) 
  • Immunogen Mouse PKA [RI] subunit aa. 225-381
  • Storage Buffer Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
  • Regulatory Status RUO
Product Details Recommended Assay References

Description

cAMP-dependent Protein Kinase (PKA) is composed of two distinct subunits: catalytic (C) and regulatory (R). Four regulatory subunits have been identified: RIα, RIß, RIIα, 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 (Cα, Cß, or Cγ). 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 Rα 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.

Format
  • Format FITC
  • Excitation Source Blue 488 nm
  • Excitation Max 494 nm
  • Emission Max 520 nm

FITC, fluorescein isothiocyanate, is a fluorochrome with a molecular weight of 389 Da. FITC is sensitive to pH changes and photobleaching. Due to nearly identical excitation and emission properties but different spillover characteristics, FITC and Alexa Fluor® 488 cannot be used simultaneously. FITC is relatively dim and should be reserved for highly expressed markers whenever possible.

Other Formats →

Suggested Companion Products

Purified Mouse Anti- PKA [RI] 18/PKA [RI] RUO
50 µg Cat No: 610165

Resources & Tools
 Spectrum Viewer  Panel Designer  Spectrum Viewer  Download TDS  Regulatory Document Website
Preparation and Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with FITC under optimum conditions, and unreacted FITC was removed. Store undiluted at -20°C.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.


Bioimaging: Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Bioimaging_Certified.shtml

Methanol Procedure for a 96 well plate:

Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 90% methanol. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Wash three times with PBS. Flick out PBS and add second step reagent. Incubate for 1 hour at RT. Wash three times with PBS. Image sample.

Triton-X 100 Procedure for a 96 well plate:

Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 0.1% Triton-X 100. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Flick out and wash three times with PBS. Flick out and add second step reagent. Incubate for 1 hour at RT. Flick out and wash three times with PBS. Image sample.


  1. Cho-Chung YS. Role of cyclic AMP receptor proteins in growth, differentiation, and suppression of malignancy: new approaches to therapy. Cancer Res. 1990; 50(22):7093-7100. View reference

  2. Dohrman DP, Diamond I, Gordon AS. Ethanol causes translocation of cAMP-dependent protein kinase catalytic subunit to the nucleus. Proc Natl Acad Sci U S A. 1996; 93(19):10217-10221. View reference

  3. Rohlff C, Clair T, Cho-Chung YS. 8-Cl-cAMP induces truncation and down-regulation of the RI alpha subunit and up-regulation of the RII beta subunit of cAMP-dependent protein kinase leading to type II holoenzyme-dependent growth inhibition and differentiation of HL-60 leukemia cells. J Biol Chem. 1993; 268(8):5774-5782. View reference

  4. Taylor SS, Buechler JA, Yonemoto W. cAMP-dependent protein kinase: framework for a diverse family of regulatory enzymes. Annu Rev Biochem. 1990; 59:971-1005. View reference

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