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Description
Research Area
Immunology & Inflammation
Images & Validation
−Item 1 of 2
| Tested Applications | ELISA, IHC, IP, WB |
|---|---|
| Dilution Range | ELISA: 1:500 - 1:2,000, IHC: 1:100 - 1:200, IP: 1:400 - 1:800, WB: 1:1,000 |
| Reactivity | Human |
| 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 -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. 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 | Liquid (sterile filtered) |
| Buffer/Preservatives | Preservative: None. Stabilizer: None; Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 |
| Concentration | 1.1 mg/mL |
| Expiration Date | 12 months from date of receipt. |
| Dry Ice Shipping | Please note: This product requires shipment on dry ice. A dry ice surcharge will apply. |
| Disclaimer | For research use only |
Alternative Names
−rabbit anti-IL-1 beta Antibody, rabbit anti-interleukin-1 beta antibody, IL-1 beta, Interleukin-1 beta, IL-1ß, catabolin
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Immunohistochemistry of Human IL1 beta antibody. Tissue: human medullary lymph node. Fixation: formalin fixed paraffin embedded. Antigen retrieval: user optimized. Primary antibody: Human IL1 beta antibody. Secondary antibody: Peroxidase goat anti-rabbit at 1:10000 for 45 min at RT. Localization: cytoplasm. Staining: Close up of medullary lymph node: positive staining in the cytoplasm of circulating macrophages. Neg Ctr (far right) of normal rabbit IgG with pH6.2 at 40X.
Western Blot of Rabbit anti-Human IL-1ß antibody. Lane 1: Human IL-1ß. Load: 5 ng per lane. Primary antibody: Human IL-1ß antibody at 1:2000 for overnight at 4°C. Secondary antibody: Peroxidase rabbit secondary antibody at 1:40000 for 30 min at RT. Block: Blocking Buffer for Fluorescent Western Blotting (orb348637) for 30 min at RT. Predicted/Observed size: 17 kDa, 17 kDa for Human IL-1ß. Other band(s): Unspecific band at ~35 kDa.
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Protocol Information
WB
Western Blot (IB, immunoblot)
IHC
Immunohistochemistry
ELISA
Enzyme-linked Immunosorbent Assay (EIA)
IP
Immunoprecipitation
IL1B Antibody (orb345121)
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