Merck, 407143-100UG, Anti-IP3 Receptor Rabbit pAb

General description
Rabbit polyclonal antibody purified by ammonium sulfate precipitation and ion-exchange (DEAE) column chromatography. Recognizes the ~260 kDa IP₃ receptor protein.
Recognizes all of the ~260 kDa IP₃ receptor subtypes.
This Anti-IP₃ Receptor Rabbit pAb is validated for use in ELISA, FC, Immunoblotting, IF, IHC, IP, Radioimmunoassay for the detection of IP₃ Receptor.
Immunogen
a synthetic peptide corresponding to amino acids in the C-terminal cytoplasmic domain of the IP₃ receptor
Application
ELISA (1:2000)
Flow Cytometry (1:1000)
Immunoblotting (1:250)
Immunofluorescence (1:200)
Immunohistochemistry (1:200)
Immunoprecipitation (1:200)
Radioimmunoassay (1:10000)
Physical form
In 150 mM NaCl, 10 mM sodium phosphate, pH 7.5.
Preparation Note
Following initial thaw, aliquot and freeze (-20°C).
Other Notes
Bourguignon, L.Y.W., et al. 1994. J. Cell Physiol. 159, 29.
Bourguignon, L.Y.W., et al. 1993. J. Biol. Chem.268, 7290.
Bourguignon, L.Y.W., et al. 1993. Cell Biol. Int.17, 751.
Variables associated with assay conditions will dictate the proper working dilution.

Recommended Protocol for Immunoblotting using anti-IP₃ Receptor Antibodies

NOTE: For protein loading, we recommend 1 x 10⁶ cells/gel lane or 100 μg/gel lane.

• TBST
• 20 mM Tris-HCl, pH 7.6
• 150 mM NaCl
• 0.05% Tween®-20 detergent

1. After transfer, block in 15 ml of TBST with 5% dry milk (for polyclonal antibody) or 1% BSA or 1-2% gelatin (for monoclonal antibody).
2. Dilute primary antibody in 5% dry milk (for polyclonal antibody) or 1% BSA or 1-2% gelatin (for monoclonal antibody) in TBST.
3. Incubate with polyclonal antibody for 1 h or with monoclonal antibody for 2 h.
4. Rinse twice rapidly, followed by 1 x 15 min and 2 x 5 min washes with TBST.
5. Dilute secondary antibody also in 5% dry milk (for polyclonal antibody) or 1% BSA or 1-2% gelatin (for monoclonal antibody) in TBST, followed by HRP.
6. Shake for 1 h.
7. Rinse twice rapidly, followed by 1 x 15 min and 2 x 5 min washes with TBST.
8. Mix development solution (1:1) (Amersham ECL kit).
9. In between two pieces of transparency add development solution. Allow to sit for approximately 1 min before exposure to film.
Legal Information
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
TWEEN is a registered trademark of Croda International PLC
Disclaimer
Toxicity: Standard Handling (A)