Technical Data

G8972-ML550
Clone Type
Monoclonal
Host
Mouse
Source
Human
Conjugate
MaxLight™550
Isotype
IgG2a,k
Clone Number
0.T.54 (C5/1.1)
Grade
Affinity Purified
Applications
IP WB
Crossreactivity
Bo Ch Hu Mo Po Rb Rt
Accession #
NM_001619, NM_005160
Shipping Temp
Blue Ice
Storage Temp
4°C Do Not Freeze
Notes
Preservative Free
BSA Free
Mouse Anti-GRK 2,3 (betaARK 1,2) (MaxLight 550)

MaxLight™550 is a new Yellow-Green photostable dye conjugate comparable to Alexa Fluor™546, 555, DyLight™549 , Cy3™, TRITC and offers better labeling efficiency, brighter imaging and increased immunodetection. Absorbance (550nm); Emission (575nm); Extinction Coefficient 150,000.

Applications
Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested.
Recommended Dilutions
Immunoprecipitation: This antibody can be used to immunoprecipitate GRK2/3 from transfected cells. Western Blot: 0.1-1ug/ml. Detects GRK2/3 in RIPA lysates from mouse 3T3 fibroblasts. Previous lots of this antibody detected GRK2/3 in bovine brain cytosol preparation, rat L6 myoblasts, human A431 carcinoma and human HeLa cell RIPA lysates. 3T3 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with G8972 (0.1ug/ml). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system.
Recommended secondary antibodies
I1904-06C: IgG, H&L (HRP) (X-Adsorbed) Pab Gt xMo I1904-06H IgG, H&L (HRP) (X-Adsorbed) Pab Gt xMo I1904-14C: IgG, Fc (HRP) Pab Gt xMo I1904-19C: IgG, F(ab’)2 (HRP) Pab Gt xMo
Positive Control Included
F4209: Fibroblast Cell, 3T3/A31. Add 2.5ul of 2-mercaptoethanol/100ul of lysate and boil for 5 minutes to reduce the preparation. Load 20ug of reduced lysate per lane for minigels.
Storage and Stability
May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Note: Applications are based on unconjugated antibody.
Immunogen
Partial GST fusion protein corresponding to the GRK3 (bARK2) carboxyl terminus (aa 467-688). This immunogen shares homology with GRK2 (bARK1). The minimal epitope recognized by the antibody is aa 483-485.
Form
Supplied as a liquid in PBS, pH 7.2. No preservative added. Labeled with MaxLight™550.
Purity
Purified by Protein G affinity chromatography.
Specificity
Recognizes human GRK2/3 at ~80 kD. Additional bands at 85 and 50 kDa are also detected when using higher concentrations of the antibody. Epitope maps to aa483-485. Species Crossreactivity: rat, mouse, chicken, bovine, porcine and rabbit.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

References
1. DebBurman, S.K., et al., J. Biol. Chem. 271: 22552-25564. 2, 1996. 2. Chuang, T.T., et al., J. Biol. Chem. 271: 28691-2869, 1996. 3. Application References: Daaka, Y., et al., Proc. Natl. Acad. Sci. USA 94: 2180-2185, 1997. 4. Freedman, N.J., et al., J. Biol. Chem. 272: 17734-17743,1997. 5. ppermann, M., et al., Proc. Natl. Acad. Sci. USA 93: 7649-7654, 1996.
USBio References
No references available
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