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Invitrogen
IL-1 beta (Pro-form) Monoclonal Antibody (NJTEN3), APC-eFluor™ 780, eBioscience™
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FIGURE: 1 / 27
IL-1 beta (Pro-form) Antibody (47-7114-80) in Flow
Mouse thioglycolate-elicited peritoneal macrophages were unstimulated (left) or stimulated for 5 hours with LPS (right) in the presence of Protein Transport Inhibitor Cocktail (Product # 00-4980-03). Cells were then surface stained with Anti-Mouse CD14 FITC (Product # 11-0141-82) followed by fixation and permeabilization with the Intracellular Fixation & Permeabilization Buffer Set (88-8824), and intracellu... View More
Product Details
47-7114-80
Applications
Tested Dilution
Publications
Product Specifications
Published species
Human,
Mouse
Host/Isotype
Rat
/ IgG1, kappa
Recommended Isotype Control
Class
Monoclonal
Type
Antibody
Clone
NJTEN3
Conjugate
Excitation/Emission Max
756/785 nm
View spectra
Form
Liquid
Concentration
0.2 mg/mL
Storage conditions
4°C, store in dark, DO NOT FREEZE!
Shipping conditions
Wet ice
RRID
AB_2573995
Product Specific Information
Description: This NJTEN3 monoclonal antibody reacts with the pro-form of mouse IL-1 beta, which is a proinflammatory cytokine expressed by monocytes, macrophages, and dendritic cells. It is synthesized in response to inflammatory stimuli as a 31 kDa inactive pro-form that accumulates in the cytosol. Cleavage of pro-IL-1 beta into the active 17 kDa protein requires the activation of inflammasomes, which are multi-protein complexes that respond to pathogens, stress conditions, and other danger signals. Inflammasome activation triggers the processing of the caspase-1 precursor into its active form, which in turn cleaves pro-IL-1 beta. IL-1 beta lacks a signal sequence peptide for classical ER/Golgi pathway and is instead secreted alongside caspase-1 via an alternate and incompletely understood mechanism. IL-1 beta signals via the IL-1RI, which is shared with IL-1 alpha. These cytokines play important roles in innate host defense by triggering the production of other proinflammatory cytokines in target cells and initiating acute-phase responses. Their activity can be moderated by IL-1 Receptor Antagonist (IL-1RA), a protein produced by many cell types that blocks receptor binding through competitive inhibition. Elevated levels of IL-1 beta have been associated with many chronic inflammatory conditions, giving IL-RA or IL-1 beta neutralizing antibodies potential therapeutical value. The NJTEN3 antibody recognizes only the pro-form of mouse IL-1 beta and does not see the active (cleaved) form.
Applications Reported: This NJTEN3 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.
Applications Tested: This NJTEN3 antibody has been tested by intracellular staining and flow cytometric analysis of stimulated mouse thioglycolate-elicited peritoneal macrophages using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Please refer to BestProtocols®: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins. This can be used at less than or equal to 0.125 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
APC-eFluor® 780 emits at 780 nm and is excited with the Red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochrome.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 633-647 nm; Emission: 780 nm; Laser: Red Laser.
Filtration: 0.2 µm post-manufacturing filtered.
Target Information
Interleukin-1 beta (IL-1 beta) is a proinflammatory cytokine expressed by monocytes, macrophages, and dendritic cells. It is synthesized in response to inflammatory stimuli as a 31 kDa inactive pro-form that accumulates in the cytosol. Cleavage of pro-IL-1 beta into the active 17 kDa protein requires the activation of inflammasomes, which are multi-protein complexes that respond to pathogens, stress conditions, and other danger signals. Inflammasome activation triggers the processing of the caspase-1 precursor into its active form, which in turn cleaves pro-IL-1 beta. IL-1 beta lacks a signal sequence peptide for classical ER/Golgi pathway and is instead secreted alongside caspase-1 via an alternate and incompletely understood mechanism. Although IL-1 beta is most often secreted in its active form, secretion of the uncleaved protein may be detectable under some biological conditions.IL-1 beta signals through two receptors, IL-1RI and IL-1RII, both of which are shared with IL-1 alpha. These cytokines play important roles in innate host defense by triggering the production of other proinflammatory cytokines in target cells and initiating acute-phase responses. Their activity can be moderated by IL-1 Receptor Antagonist (IL-1RA), a protein produced by many cell types that blocks receptor binding through competitive inhibition. Elevated levels of IL-1 beta have been associated with many chronic inflammatory conditions, giving IL-RA or IL-1 beta neutralizing antibodies potential therapeutical value.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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