rev: September 15, 2004
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ANTIBODIES
(anti-Human and others as indicated)
RDI Divison of Fitzgerald Industries Intl offers a wide line of antibodies. Since no one antibody works best for all applications (neutralization, blotting, histochemistry, ELISA, etc), we offer many different types of antibodies to help solve this problem. Please inquire for other applications or types of antibodies not listed below.
Rabbit anti-Rat Sodium Channel (RI and RII)
Background: Anti-Rat subtypes I and II (see ref) Na channel antibodies were
elicited from specific peptides. These subtypes are expressed differentially
in specific regions of the brain.
cat# RDI-NACHAN9Aabr $438.00 antigen = SP1
(19 amino acids)
RDI-NACHAN9Babr $438.00 antigen = SP11I (18 amino acids)
RDI-NACHAN9Cabr $438.00
antigen = SP11II (20 amino acids)
LOT# see sheet with shipment
Packaging: 1 milligram IgG fraction in 1ml 0.1M Tris-Hcl, pH 7.5.
Species: rabbit IgG purified by Protein G Sepharose chromatography.
Antigen: each of the above antibodies were made individually with a synthetic
peptide, designated A, B and C (respectively).
Specificity: specific for the type I and Type II sodium channel
(immunoprecipitation). Cross reactivity with other Na Channels have been
observed with other mammalian species.
Uses: Optimal dilutions must be determined by the end user for their particular
application.
ref: -Gordon, D. et al, Proc. Natl. Acad. Sci. USA, 84, 8682-8686 (1987)
Storage: Store at -20 DEG C. AVOID FREQUENT FREEZE THAW CYCLES.
background:The sodium channel is comprised of a large alpha subunit and two smaller Beta subunits. The Type I NA channels are expressed in the brain stem, cortex, substantia nigra and tectum of the adult brain, while the type II Na Cahnnels alpha subunits are expressed in myelinated axons of the adult mammalian brain. The Type III Na Channels alpha subunit is expressed in embryonic, but not adult brain. The Na channel protein contains four internal repeats (S1,S2,S3,S5,S6) and one positively charged segment (S4) thought to be the voltage-sensor.
cat# RDI-NACHAN2abm $594.00/vial
Packaging: 200 micrograms ammonium sulfate precipiatted and DEAE chroimatography
purified IgG in 200ul 0.1M Tris, pH 7.5, with 30% glycerol and 0.05% sodium
azide
Species: mouse IgG1
Clone ref: K69/3
Antigen: fusion protein corresponing to residues 1882- 2005 of rat brain
soidum channel protein type II, a subunit.
Specificity: specific for the type II sodium channel alpha- subunit (Mr
220-260kDa)
Species Reactivity: Rat, Mouse and Human (others not tested)
Uses: -western blot: 0.5-2ug/ml to detect the sodium channel type II from
rat brain microsomal prep (4 DEG C overnight and using enhanced chemiluminescence
detection , bands 220- 260kDa).
Optimal dilutions must be determined by the end user for their particular
application.
General ref: -Gong, B et al, Soc. Neurosci Abstr,; 23:909,1997
Storage: Store at -20 DEG C. AVOID FREQUENT FREEZE THAW CYCLES. Upon Receipt,
centrifuge for 1 minutes at 100rpm to concentarte antibody in vial
For Research Use Only-Not for Use in Diagnostics
-copyright by owner
Immunoblot Protocol
1. Perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on a microsome preparation and transfer the proteins to nitrocellulose. Do not boil microsomes. Warm microsomes to 37°C for 30-45 minutes before loading the sample. Wash the blotted nitrocellulose twice with water.
2. Block the blotted nitrocellulose in freshly prepared PBS containing 3% nonfat dry milk and 0.05% Tween-20 (PBST-MLK) for 30 minutes at 20-25°C with constant agitation.
3. Incubate the nitrocellulose with 0.5-2mg/ml of anti-Na + Channel, Type ll, diluted in freshly prepared PBST-MLK overnight with agitation at 4°C.
4. Wash the nitrocellulose twice with water.
5. Incubate the nitrocellulose in the secondary reagent of choice (a goat anti-mouse HRP conjugated IgG, (1:5000 dilution was used) in PBST-MLK for 1.5 hours at room temperature with agitation.
6. Wash the nitrocellulose with water twice.
7. Wash the nitrocellulose in PBS-1.0 % Tween 20 for 3-5 minutes.
8. Rinse the nitrocellulose in 4-5 changes of water.
9. Use detection method of choice (enhanced chemiluminescence was used).
cat#RDI-NACHAN3abrX $562.00/200ug in Tris glycine/glycerol
-antigen: peptide designated EOIII (TEEQKKYYNAMKKLGSKKC) corresponding to the intracellular III-IV loop (identical in all known veterbrates)
-sutiable for : western blot (1-2ug/ml) 220Kda using rat brain micrsome extract, immunoprecipitation (1-4ug/ml) cytochemistry 20ug/ml (mouse sciatic nerves as positive control)
-expected to react with all known veterbrates
34 Junction Square Drive
Concord MA 01742-3049
USA
phone (978) 371-6446 or (800) 370-2222
fax (978) 371-2266
EMAIL:antibodies@fitzgerald-fii.com
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