Analysis Note

ControlPositive Control: Cerebral cortex and pyramidal neurons, mouse brain lysate.

Application

Anti-Glutamate Receptor 2 Antibody, extracellular, clone 6C4 detects level of Glutamate Receptor 2 & has been published & validated for use in ELISA, IC, IH, IP, RIA & WB with more than 50 product citations.

Immunocytochemistry:
on 4% paraformaldehyde fixed cells was used in a previous lot:2-3 µg/mL (Vissavajjhala, 1996; Osten, 1998; Passafaro, 2001).

ELISA/RIA:
A previous lot of this antibody was used in ELISA/RIA.(Vissavajjhala, 1996).

Immunohistochemistry on 50 µm, 4% paraformaldehyde fixed brain sections: 1:500-1:800 (Vissavajjhala, 1996; Yung, 1998; Kumar; 2002).

Immunoprecipitation: 2-4 µg/mL (Osten, 1998).

Western blot: 1-2 µg/mL (Vissavajjhala, 1996; Osten, 1998); membrane preparations are suggested for enhanced signals.

Optimal working dilutions and protocols must be determined by end user.

General description

Glutamate receptors (GluRs) can be categorized as ionotropic or metabotropic and subcatergorized by their agonist preferences (NMDA, AMPA or Kainic acid). There are four types of AMPA selective GluR subunits (GluR1, GluR2, GluR3 and GluR4). Tetrameric or pentameric combinations of different subunits contributes to the functional diversity of AMPA receptors. In general, AMPA receptors mediate fast synaptic current at most excitatory synapses, with stoichiometry characterized by subtype composition. Although subunit composition of AMPA receptors varies, they must contain at least one edited GluR2 subunit to be calcium impermeable. The critical residue controlling calcium permeability is in the pore loop region. In GluR1, GluR3, and GluR4, this positionis occupied by a Gln residue. In GluR2, it is occupied by an Arg residue. It has been shown experimentally that the presence of Arg in this position blocks CA2+ ion permeability, while a Gln does not. Relative calcium permeability in AMPA receptor channels may be significant in pathological neurotoxic damage and long term changes in nervous system responses.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

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

Physical form

Purified immunoglobulin from culture supernatant

Format: Purified

Purified mouse immunoglobin IgG2a liquid in buffer containing PBS, no preservative.

Quality

Routinely evaluated by Western Blot on mouse brain lysates.

Western Blot Analysis:
1:1000 dilution of this lot detected glutamate receptor 2 on 10 µg of mouse brain lysates.

Specificity

Recognizes the large N-terminal extracellular domain of Glutamate Receptor 2 (GluR2). No cross-reactivity observed with other AMPA/Kainate GluR subunits. On western blots of brain extracts from rat, macaque monkey, and dog, MAB397 recognizes a band at approximately 102 kDa corresponding to full length GluR2. Other proteins noted by Western blot at 66 kDa or lower molecular weight appear to be breakdown products of GluR2 (Vissavajjhala, 1996). By immunohistochemistry GluR2 is widely distributed at both the cellular and synaptic levels. MAB397 recognizes GluR2 present in a vast majority of, but not all, GABAergic interneurons (Vissavajjhala, 1996).

Storage and Stability

Stable for 6 months at -20°C in undiluted aliquots from date of receipt.
Handling Recommendations: Upon receipt, 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 the IgG2a and affect product performance.

Target description

102 kDa