Analysis Note

ControlRAW264 cell lysate

Application

Immunohistochemisty (paraffin): IP3 Receptor staining in rat cerebellum, tissue was pretreated with citrate buffer, pH 6.0. A previous lot of this antibody was diluted to 1:100, using IHC-Select™ detection with HRP-DAB.

Immunofluorescence: 1:20 - 1:200 dilution from a previous lot. Light fixation, 2% PFA, 90% ethanol fixation; Visualization by confocal microscopy is required, as detection by standard fluorescent microscopy will not be adequate to detect the IP3R. Due to the intensity of confocal lasers, use of an anti-fading agent, such as DABCO, is strongly recommended.

ELISA: 1:2,000 dilution of a previous was used in ELISA.

Immunoprecipitation: 1:200 dilution of a previous lot was used in Immunoprecipitation.

Flow cytometry: 1:1,000 dilution of a previous lot was used in Flow Cytometry.

Radioimmunoassay: 1:10,000 dilution of a previous lot was used in Radioimmunoassay.

Optimal working dilutions must be determined by the end user.

Research CategorySignaling

Research Sub CategoryGPCR, cAMP/cGMP & Calcium Signaling

This Anti-IP3 Receptor Antibody is validated for use in WB, IF, IH(P), ELISA, IP, RIA for the detection of IP3 Receptor.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

The InsP3 (inositol 1,4,5-trisphosphate) Receptor family are intracellular Ca2+ channels activated by the second messenger InsP3. There are at least three types of receptor, with distinct tissue-specific expression profiles and subcellular localizations; it is thought that the exact subtype profile conveys precise Ca2+ release responses. Release of Ca2+ from intracellular stores is the trigger for many cellular responses, including muscle contraction. Receptor activity is subject to feedback regulation by Ca2+ concentration, as high cytosolic Ca2+ concentrations shut down the channel. Ca2+ release is augmented by phosphorylation of the Receptor by PKA and PKG.

Immunogen

Synthetic peptide (KDSTEYTGPESYV) corresponding to the C-terminus of human IP3 Receptor type 1.

Epitope: C-terminus

Linkage

Replaces: AB1622

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Purified

Format: Purified

Purified rabbit polyclonal IgG in buffer containing 10 mM sodium phosphate, 150 mM NaCl, and 0.01% sodium azide, pH 7.5.

Quality

Routinely evaluated by Western Blot on RAW 264 lysates.

Western Blot Analysis:
1:500-1,000 dilution of this lot detected IP3 Receptor on 10 µg of RAW 264 lysates.

Specificity

Reacts with the C-terminal cytoplasmic domain of IP3 receptors Type 1, 2, and 3. IP3 receptors have been shown to play an important role in the regulation of Ca2+ release from internal storage sites during cell activation. AB1622 reacts with IP3 receptor (MW approx. 260 kDa) isolated from a variety of cell types (e.g. lymphocytes, macrophages, granulocytes, fibroblasts, epithelial, endothelial cells, skeletal muscle, cardiac muscle and brain tissues).

Reactivity with other species has not been determined.

Storage and Stability

Stable for 1 year at -20°C in undiluted aliquots from date of receipt.
Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

~260 kDa