rev:  February 10, 2004

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INSULIN  RELATED 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.


DATA SHEET: Mouse anti-Human Insulin Like Growth Factor I (II*)

Catalog#: RDI-ILGF12abm       Price: $562.00/vial      

Package Size: 200 micrograms lyophilized 308ul of 0.1M Tris-glycine pH7.4, 0.15M NaCl, 0.1M EDTA. 0.2micron filtered. no preservatives or carrier added) Reconstitute with 500ul sterile water (yielding a 400ug/ml stock solution)

Species: mouse IgG1 Protein G purified

Antigen: purified human IGF-I from human plasma

Specificity: specific for IGF-I of human, rat and mouse origin (**approx 40% cross with ILGF II), some cross with chicken IGF-I & 2. UPDATED CROSS REACTIVITY Study see Endocrinology, Oct 1997

Activity: Neutralization: 10-20 micrograms/ml to inhibit the stimulatory effect of 10 nanograms IGF-I/ml in cultures of chicken embryo fibroblasts.(neutralization can be hindered by stearic effects of rat IGF-1 binding protein.

Western blot: 0.5-1 micrograms/ml to detect 100ng IGF-I/IGF-II under non reducing conditions. Recommend overnight 4 DEG C incubation with enhanced chemiluminescence detection

-MUST TITER FOR EACH APPLICATION

-Immunoprecipitation (approx 5ug for 100ng IGF-1 using Protien-G Agarose)

-Immunohistochemistry (approx 10ug/ml for paraffin embedded methanol:acetone 50:50 fixed skin sections)

Storage: Upon receipt, store at -20 DEG C or below. -After reconstitution, aliquot and store at - 20 DEG C. Avoid frequent freeze thaw cycles.

ref: -Van Wyk, J.J. et al, Endocrinology Oct 1997

       -Russell, et al, Proc. Natl. Acad Sci 81:2389-2392(1984)

      -Gillespie, G.Y. et al, Meth. Enz. 146:207   (1987)

       -Van Wyk, J.J. et al, Human Growth Hormone,   585-599. Plenum Publishing Corp, NY (1986)

Precautions: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 products.


NOTE: When attempting to use a monoclonal antibody against rat or mouse proteins, special precautions must be taken to limit non specific binding of your detecting antibody and the blotted mouse proteins. This activity is due to the requirement that your detecting antibody bind to the mouse primary antibody but by its nature, it will also bind to many other mouse proteins!


1) For rat, use a second antibody that has been affinity absorbed against rat proteins, ie, goat anti-mouse IgG:HRP absorbed against rat, human proteins. etc

2) For mouse, you may need to try the following:   a) directly label the detecting specific antibody, ie biotin

label or HRP label the mouse anti-ILGF1 ab (obtain conjugation kit fromm Pierce Biochemical or Sigma chemical).


-OR-

b) use a subclass specific detecting ab, ie, goat anti-mouse IgG1:HRP or goat anti-mouse IgG1:Biotin. This can help eliminate some of the seconday antibodies binding to the blotted proteins non specifically.

-And/or-


c) Running the blot as usual, try using a few dilutions of the detecting ab (ECL system etc) without using any of the primary antibody. Use the most concentrated dilution of the labeled second antibody that gives you the least activity, ie signal. This will limit your overall sensitivity, but may reduce much of the non specific signal.



click here for our specialty second antibodies

RDI Divison of Fitzgerald Industries Intl

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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