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Description
Research Area
Images & Validation
−| Tested Applications | ELISA, WB |
|---|---|
| Dilution Range | ELISA: 1:1,000 - 1:5,000, WB: 1:500 - 1:2,000 |
| Reactivity | Canine, Human, Primate |
| Application Notes |
Key Properties
−| Antibody Type | Primary Antibody |
|---|---|
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Immunogen | This antibody was prepared by repeated immunizations with recombinant human IL-1ß produced in E.coli. The MW of the recombinant 153 aa IL-1ß was 17 kDa with the N-terminal amino acid at position alanine 117. This cleavage site is generated by the IL-1ß converting enzyme (ICE, capase-1). |
| Target | IL1B |
| Purity | This is an IgG preparation of whole rabbit serum purified by DEAE fractionation. This antibody is primarily directed against mature, 17,000 MW human IL-1ß and is useful in determining its presence in various assays. In general, this antibody also detects primate IL-1ß in the same formats using similar dilutions. The antiserum does not recognize human IL-1α. In ELISA formats and other immunoreactive assays, this antibody will recognize 10% of the non-denatured (native) precursor 31,000 MW IL-1ß containing samples but will primarily detect all of the 17,000 MW mature molecule. However, in immunoblot analysis of natural cell products or human body fluids, the usual procedure of hearing the sample in SDS with or without reducing agents will facilitate denaturing of the 31,000 MW IL- 1ß precursor molecule. Denatured 31,000 precursor IL-1ß will be recognized by this antibody but often migrates as a 35,000 MW band. This is due to the unfolding of the denatured precursor IL-1ß exposing epitopes not exposed in the natural state. In immunoblots, depending on the number of cells, the antibody detects the 17,000 MW band in supernatants as well as a 35,000 MW band representing the 31,000 MW IL-1ß precursor in lysates. |
| Conjugation | Unconjugated |
Storage & Handling
−| Storage | Store vial at 4° C prior to restoration. For extended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use. |
|---|---|
| Form/Appearance | Lyophilized |
| Buffer/Preservatives | Preservative: None. Stabilizer: None; Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 |
| Concentration | 1.0 mg/mL |
| Expiration Date | 12 months from date of receipt. |
| Disclaimer | For research use only |
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Western blot showing detection of recombinant human IL-1ß by Biorbyt's anti-IL-1ß antibody. Recombinant human IL-1ß was loaded on to an SDS-PAGE gel at 0.1 µg and after separation, transferred to nitrocellulose. The expected band is approximately 18 kDa in size. The membrane was blocked with 1% BSA in TBST for 30 min at RT, followed by incubation with primary antibody diluted 1:1000 in 1% BSA in TBST overnight at 4°C. After washes, the blot was reacted with secondary antibody HRP Goat anti-Rabbit IgG antibody diluted 1:40000 in blocking buffer (p/n orb348637) for 30 min at RT.
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IL1B Antibody (orb345138)
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