IL-1 beta (Pro-form) Monoclonal Antibody (NJTEN3), PerCP-eFluor 710, eBioscience

PRODUCT DETAILS

Host: Rat

Isotype: IgG1, kappa

Clonality: Monoclonal

Clone: NJTEN3

Format: PerCP-eFluor 710

Reactivity: Ms

Application: Flow Cytometry

Tested Dilution: 0.25 µg/test

Concentration: 0.2 mg/mL

Storage: 4°C, store in dark, DO NOT FREEZE!

Formulation: PBS with 0.09% sodium azide; pH 7.2

Purification: Affinity chromatography

Data Sheet: TDS

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 0.25 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.

PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-822-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) 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: 488 nm; Emission: 710 nm; Laser: Blue Laser.

Filtration: 0.2 µm post-manufacturing filtered.

 

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.