rev: February 7, 2005
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ANTIBODIES
(anti-Human and others as indicated)
Research Diagnostics Inc 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.
see also anti-epo (erythropoietin)
Rabbit anti- Anti-Epo Receptor (Polyclonal) Human/Mouse
cat# RDI-EPORE1abrX (Discontinued) $475.00
Presentation: 200ug of immunoaffinity purified rabbit IgG in 0.2ml bufer of a 70:30 mix of 0.1M Tris- glycine, pH7.4, 0.15M NaCl,0.05% sodium azide and 30% glycerol. Liquid at -20 C.
Immunogen: KLH-conjugated peptide (APPPNLPDPKFESKA-C) corresponding to amino acids 25-39 of the Human EPO receptor precursor (accesion#NP_000112.1)
Specificity: Recognizes Epo Receptor, Mr 64-70kDa.
A doublet is often observed, representing the native and glycosylated forms of the receptor. Additional unknown protein at 58 is also observed in TF-1 (band inhibited when antibody is preincubated by immunizing peptide)
Species reactivity: Mouse and human. Other species not tested.
Uses: -Immunoblot Analysis: 1-2ug/ml of this lot detected Epo Receptor in RIPA lysates from TF-1 and NFC-MEL cells.
-not tested for other applicaitons
Storage: Stable for 2 years at -20°C from date of shipment. For maximum recovery of product, centrifuge the vial prior to removing the cap.
FOR RESEARCH USE ONLY NOT FOR USE IN HUMANS
Quality Control Testing Additional Research Applications
General Reference: 1. Migliaccio, A.R., et al., Proc. Natl. Acad. Sci. 88: 11086-11090, 1991.
Immunoblot Protocol
1. Perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on a cell lysate sample (cell lysis buffer: 50mM Tris-HCl, pH 7.4; 1% NP-40; 0.25% sodium deoxycholate; 150mM NaCl; 1mM EGTA; 1mM PMSF; 1 ug/ml each aprotinin, leupeptin, pepstatin; 1mM Na3VO4; 1mM NaF) and transfer the proteins to nitrocellulose. Wash the blotted nitrocellulose twice with water.
2. Block the blotted nitrocellulose in freshly prepared 5% nonfat dry milk in TBS (TBS-MLK) with 0.05% Tween(tm)-20 for 1-2 hours at room temperature with constant agitation.
3. Incubate the nitrocellulose with 1-2ug/ml of anti-Epo Receptor, diluted in freshly prepared TBS with 5% nonfat dry milk (TBS-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-rabbit HRP conjugated IgG, in TBS-MLK for 1.5 hour at room temperature with agitation.
6. Wash the nitrocellulose with water twice.
7. Wash the nitrocellulose in TBS-0.05% 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).
For In vitro research Use Only. Not for use in or on humans or animals or for diagnostics.It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. We are not responsible for any patent infringements that might result with the use of or derivation of this product.
Pleasant Hill Road
Flanders NJ 07836
phone (800) 631-9384
or (973) 584-7093
fax (973) 584-0210
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