Anti-CaMPARI-Red [4F6]

Tool Details

*FOR RESEARCH USE ONLY

• Name: Anti-CaMPARI-Red [4F6]
• Class: Monoclonal
• Conjugation: Unconjugated
• Host: Mouse
• Application: IHC ; WB
• Description: CaMPARI (Calcium Modulated Photoactivatable Ratiometric Integrator) is a photoconvertible protein construct that allows imaging of the integrated calcium activity of populations of cells over time. CaMPARI 2.0 is the second generation of CaMPARI molecules with improve green brightness by 50% and red brightness by 250%. Additionally, CaMPARI 2.0 has faster kinetics, and a lower photoconversion rate in low calcium conditions resulting in a net 100-fold difference in green-to-red switching in low- vs. high-calcium conditions. Lastly, these different CaMPARI proteins display a range of sensitivity in their calcium binding (Kd’s ranging from 100 nM to 1 uM of free calcium). The variable domain region of the original mouse IgG1 antibody was reformatted to generate the Fab fragment and the chimeric rabbit IgG.
• Immunogen: Purified and photoconverted EosFP protein, subjected it to proteolysis, and further purified the proteolytic fragment containing the red chromophore which is identical to the equivalent region of CaMPARI1 and CaMPARI2.
• Isotype: IgG2a

Target Details

• Target: CaMPARI1 and CaMPARI2
• Target background: CaMPARI (Calcium Modulated Photoactivatable Ratiometric Integrator) is a photoconvertible protein construct that allows imaging of the integrated calcium activity of populations of cells over time. CaMPARI 2.0 is the second generation of CaMPARI molecules with improve green brightness by 50% and red brightness by 250%. Additionally, CaMPARI 2.0 has faster kinetics, and a lower photoconversion rate in low calcium conditions resulting in a net 100-fold difference in green-to-red switching in low- vs. high-calcium conditions. Lastly, these different CaMPARI proteins display a range of sensitivity in their calcium binding (Kd’s ranging from 100 nM to 1 uM of free calcium).

Applications

• Application: IHC ; WB
• Application notes: 4F6 antibody is recommended for the detection and analysis of CaMPARI via immunohistochemistry and Western blot. For instance, it was successfully used in the immunohistochemical identification of CaMPARI-Red cells in brain tissue, distinguishing them from the cells in which CaMPARI stayed in ints green, unconverted form (Ebner et al., 2019).

Handling

• Format: Liquid
• Concentration: 0.9-1.1mg/ml
• Unit size: 100 ug
• Storage buffer: PBS with 0.02% azide
• Storage conditions: -80° C
• Shipping conditions: Shipping at 4° C

References

• Moeyaert et al. 2018. Nat Commun. 9(1):4440. PMID: 30361563.