Glutamate is the main excitatory neurotransmitter in the brain.
To date five glutamate Transporters have been cloned: GLAST (EAAT1), GLT1 (EAAT2), EAAC1 (EAAT3), EAAT4, and EAAT5.
These transporters are believed to be critical in reducing potentially toxic extracellular concentration of glutamate by rapid uptake into nerve terminals and glial cells.
Synaptic vesicles in the nerve terminals play a critical role in neurotransmission.
Synaptic vesicles in the nerve terminals play a critical role in neurotransmission.
Glutamergic neurotransmission occurs through an exocytotic process involving the interaction of glutamate containing synaptic vesicles with the plasma membranes of the presynaptic ending.
An electro-chemical proton gradient generated by a V-type H+-ATPase (vacuolar-type proton-translocating ATPase) in the synaptic vesicles membrane provides the driving force for glutamate uptake.
Recently a protein, termed inhibitory protein factor (IPF), has been isolated from brain cytosol that inhibits glutamate and GABA uptake into synaptic vesicles (IC50 ~25 nM).
IPF does not inhibit ATP-independent uptake, norepinephrine uptake into chromaffin vesicles, and Na-dependent glutamate uptake into synaptosomes.
IPF refers to a three distinct proteins with ~mol wt of 138kDa (IPF-alpha), 135kD (IPF-beta), and 132kD (IPF-gamma).
IPF-a is derived from a ubiquitous, non-erythroid brain spectrin called alpha-Fodrin, a well-characterized protein previously implicated in such diverse activities as exocytosis/endocytosis, apoptosis, and NMDA-receptor activation.
However, a-Fodrin itself has no effect on glutamate uptake.
The N-terminal 1-20 aa of IPF-a, IPF-b, and IPF-g are identical with 26-45 aa of a-Fodrin (mol wt ~240kD).
Therefore, it appears that some identified protease(s) may generate IPF-a from a-Fodrin.
Applications:Suitable for use in ELISA, Western Blotting.
Other applications not tested.
Recommended Dilution:Western Blot: 1:1000-1:5000 for antiserum and 1-10ug/ml for affinity purified IgG using Chemiluminescence technique.
IPFs are approx.
~135-140kD.
ELISA: Control peptide can be used to coat ELISA plates at 1ug/ml and detected with antibodies (1:10-50K for antiserum serum and 0.5-1ug/ml for affinity pure IgG).
Optimal dilutions to be determined by the researcher.
Control Peptide: G4000-72Control peptide, because of its low MW (<3kD), is not suitable for Western.
It should be used for ELISA or antibody blocking (use 5-10 ug per 1 ul of antiserum or 1 ug of aff pure IgG) to confirm antibody specificity.
Storage and Stability:May be stored at 4°C for short-term only.
For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%).
Freeze at -20°C.
Aliquots are 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.
仕様
Size：100ul
Host：rabbit
Source Antibody：human
Grade：Serum
Purity：Purified using the control peptide-Sepharose
Form：Supplied as a liquid, neat serum, 40% glycerol.
Specificity：IPF has not yet been cloned. Its sequence is derived from the N-terminal sequencing of the purified IPF (1). Antibody crossreactivity in various species is not established.
Isotype：IgG
Calc Applications Abbrev：E WB
Calc Crossreactivity：Hu
Immunogen：A 10-aa peptide mapping at the N-terminus of human IPF (1) was synthesized, conjugated to KLH
