| This antibody is primarily directed against mature, 17kD human IL-1b and is useful in determining its presence in various assays. In general, this antibody also detects primate IL-1b in the same formats using similar dilutions. The antiserum does not recognize human IL-1a. In ELISA formats and other immunoreactive assays, this antibody will recognize 10% of the non-denatured (native) precursor 31kD IL-1b containing samples but will primarily detect all of the 17kD mature molecule. However, in Western Blot analysis of natural cell products or human body fluids, the usual procedure of heating the sample in SDS with or without reducing agents will facilitate denaturing of the 31kD IL-1b precursor molecule. Denatured 31kD precursor IL-1b will be recognized by this antibody but often migrates as a 35kD band. This is due to the unfolding of the denatured precursor IL-1b exposing epitopes not exposed in the natural state. |
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| Catalog # | I7663-11P |
| Applications | Suitable for use in Neutralization, ELISA, Radioimmunoassays, Flow Cytometry, FACS, Immunohistochemistry, Western Blot and Immunoprecipitation. Other applications not tested. |
| Recommended Dilutions | Western Blot: 1:100-1:200. To detect IL-1b from supernatants or lysates of 2x10e6 endotoxin-stimulated human peripheral blood mononuclear cells (PBMC). PBMC are stimulated for 24 hours with 1% human serum plus 10ng/ml E. coli LPS. |
| Immunoprecipitation: 1:400-1:800. Preclear the preparation with a non-specific rabbit IgG (I1904-43A) to reduce background. |
| Immunohistochemistry (Paraffin or frozen): 1:100-1:200 . |
| ELISA: 1:500-1:2000. Suggested secondary antibody: I1904-46Q, IgG, H&L (HRP). |
| RIA: 1:5000-1:10,000 |
| Neutralization (Primate/human): 1:100 for at least 4 hours before analysis. |
| Optimal dilutions to be determined by the researcher. |
| Storage and Stability | May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile 40- 50% glycerol, aliquot and store 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. |
| CAS Number | n/a |
| Clone Type | Polyclonal |
| Isotype | IgG |
| Host | Rabbit |
| Source | Human |
| Concentration | ~1mg/ml |
| Form | Supplied as a liquid in PBS, pH 7.2, 0.01% sodium azide. No stabilizing proteins added. |
| Purity | Purified by DEAE fractionation. Endotoxin: <10pg/ul. |
| Immunogen | Recombinant 153aa human IL-1b produced in E. coli. |
| Specificity | Recognizes the 17kD mature human IL-1b. |
| Species Crossreactivity | Primate. Does not neutralize IL-1a or mouse, rat or rabbit IL-1b. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
