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Invitrogen™ IL-33R (ST2) Monoclonal Antibody (RMST2-2), Super Bright™ 645, eBioscience™
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Catalog No. 64933582
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64-933-582 100 μg
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Catalog No. 64-933-582 Supplier Invitrogen™ Supplier No. 64933582
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Rat Monoclonal Antibody

Description: The RMST2-2 monoclonal antibody reacts with ST2, also known as IL-33 receptor (IL-33R) or interleukin 1 receptor-like 1. ST2 is a member of the IL-1 receptor family. In the membrane-bound form, it consists of three extra-cellular immunoglobulin domains and an intracellular toll-interleukin-1 receptor domain. ST2 forms a dimer with IL-1R accessory protein in a ligand-dependent manner. The IL-33/ST2 interaction induces the production of TH2 cytokines. ST2 also has a soluble isoform lacking transmembrane and intracellular toll-interleukin-1 receptor domains. It is believed that the soluble form functions as a decoy receptor that can block membrane bound IL-33/ST2 interaction. ST2 is expressed by TH2 lymphocytes, mast cells, eosinophils, basophils, innate lymphocytes, smooth muscle cells, and endothelial cells and is involved in host defense, allergy, and inflammation. The IL-33/ST2 interaction has also been shown to be atheroprotective. Applications Reported: This RMST2-2 antibody has been reported for use in flow cytometric analysis. Applications Tested: This RMST2-2 antibody has been tested by flow cytometric analysis. This may be used at less than or equal to 1.0 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 i...

IL-33R, also known as ST2, is a receptor for IL-33. It is a member of the IL-1 receptor family of innate receptors. Its extracellular domain consists of 3 Ig-like domains that directly interact with its co-receptor IL-1RAcP, and the ligand alarmin, IL-33. IL-33 ligation results in the association of IL-33R complex with its adaptor proteins MyD88, IRAK1, IRAK4 and TRAF6, and activation of the NF-kB, and MAPK pathways. This, in turn, instigates the release of chemokines and cytokines such as IL-5, IL-6, IL-8, and IL-13. Two most common isoforms of IL-33R that result from alternative splicing are ST2L - the membrane bound form, and ST2S - the soluble form. The membrane bound form can be released from the cell surface by proteolytic cleavage. The soluble forms have been suggested to act as decoys dampening the IL-33 signaling. IL-33R is primarily expressed by mast cells, basophils, and eosinophils. In mouse, IL-33R expression has been shown on ILC2 and Th2 cells. IL-33R signaling has been implicated in the number of immune conditions, including allergies, arthritis, atherosclerosis and sepsis.
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Specifications

Antigen IL-33R (ST2)
Applications Flow Cytometry
Classification Monoclonal
Clone RMST2-2
Concentration 0.2 mg/mL
Conjugate Super Bright 645
Formulation PBS with BSA and 0.09% sodium azide; pH 7.2
Gene IL1RL1
Gene Accession No. P14719
Gene Alias DER4; Fit1; Fit-1; Fos-responsive gene 1 protein; growth stimulation-expressed; homolog of mouse growth stimulation-expressed; IL-1 R4; IL1RL1; IL-1RL1; IL33R; IL-33R; interleukin 1 receptor like 1; interleukin 1 receptor-like 1; interleukin 1 receptor-related protein; Interleukin1 receptor like 1; interleukin-1 receptor-like 1; Interleukin-33 receptor alpha chain; Ly84; lymphocyte antigen 84; Protein ST2; Protein T1; sIL 33R; soluble IL 33 Receptor; soluble IL 33R; St2; ST2L; St2-rs1; ST2V; Ste2; T1; T1/ST2
Gene Symbols IL1RL1
Host Species Rat
Purification Method Affinity chromatography
Quantity 100 μg
Regulatory Status RUO
Primary or Secondary Primary
Gene ID (Entrez) 17082
Target Species Mouse
Content And Storage 4°C, store in dark, DO NOT FREEZE!
Product Type Antibody
Form Liquid
Isotype IgG2a κ
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Can I use the OneComp and UltraComp eBeads microspheres with Super Bright-conjugated antibodies?

UltraComp eBeads microspheres (Cat. No. 01-2222) are recommended for use with Super Bright dyes.
Note: Super Bright Staining Buffer (Cat. No. SB-4400) is not compatible with UltraComp eBeads microspheres (Cat. No. 01-2222-41, 00-2222-42). If using UltraComp eBeads microspheres as a compensation tool, solely use Flow Cytometry Stain Buffer (Cat. No. 00-4222-26, 00-4222-57) for any antibody dilutions.

In some experiments, we have observed that compensation values for Super Bright 780- and Brilliant Violet 785- or Brilliant Violet 786-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads, both with Super Bright 780 and Brilliant Violet 786. We have not tested Brilliant Violet 785 with the AbC beads.

Can I prepare an antibody cocktail containing Super Bright Staining Buffer and Super Bright-conjugated antibodies ahead of time?

We recommend that the antibody cocktails containing Super Bright-conjugated antibodies and Super Bright Staining Buffer are prepared fresh prior to staining. Discard any unused portions. We do not recommend overnight storage of prepared cocktails.

Can I fix my cells after staining with Super Bright-conjugated antibodies? How long can fixed cells be stored prior to analysis?

Samples that have been stained with antibodies conjugated to Super Bright dyes may be stored for up to three days, at 2-8°C, in the dark, using either IC Fixation Buffer (Cat. No. 00-8222) or 1-step Fix/Lyse Buffer (Cat. No. 00-5333) with no significant effect on brightness or compensation.

Are the Super Bright dyes sensitive to methanol fixation?

Super Bright dyes are stable in methanol-based fixation buffers.

Can Super Bright-conjugated antibodies be used in combination with intracellular (IC Fixation/Permeabilization) or intranuclear (Foxp3 fixation/permeabilization) staining?

Yes, Super Bright-conjugated antibodies are stable in formaldehyde-based fixation buffers and permeabilization buffers, such as the IC Fixation and Permeabilization Buffer Set (Cat. No. 88-8824) and the Foxp3/Transcription Factor Staining Buffer Set (Cat. No. 00-5523).

Which laser do I use to detect Super Bright-conjugated antibodies?

The violet laser (405 nm) should be used to excite Super Bright-conjugated antibodies.

Can the Super Bright Staining Buffer be used with other polymer dyes?

Yes, the Super Bright Staining Buffer (Cat. No. SB-4400) is compatible with other polymer dyes (i.e., Brilliant Violet dyes) and is useful for minimizing any non-specific polymer interactions when two or more of these dyes are used in combination.

Can Super Bright-conjugated antibodies be used in combination with other polymer dyes? What buffer should I use when using more than one Super Bright or polymer dye?

When using two or more Super Bright dyes, or when using Super Bright dyes in combination with other polymer dyes (i.e., Brilliant Violet dyes), we recommend using the Super Bright Staining Buffer (Cat. No. SB-4400) to minimize any non-specific polymer interactions between these fluorochromes. For specific instructions for use, please refer to the product technical data sheet.

Are there recommended tubes for staining with Super Bright-conjugated antibodies?

We recommend using polystyrene tubes (for example, FACS tubes) for staining with your Super Bright-conjugated antibodies. If using polypropylene (for example, Eppendorf tubes), protecting from light is critical.

Is there a specific buffer I should use when staining with Super Bright-conjugated antibodies?

No special buffer is required when using a single Super Bright-conjugated antibody in a panel.
When using more than one Super Bright dye, or when using Super Bright dyes in combination with other polymer dyes (i.e., Brilliant Violet dyes), we recommend using the Super Bright Staining Buffer (Cat. No. SB-4400) to minimize any non-specific polymer interactions between these fluorochromes. For specific instructions for use, please refer to the product technical data sheet.

How does the protocol for staining with Super Bright-conjugated antibodies differ from staining with conventional organic fluorochrome and eVolve conjugated antibodies?

Antibodies conjugated to Super Bright dyes can be used similarly to traditional fluorochromes. If multiple Super Bright dyes are used in combination with each other or in combination with other polymer dyes (i.e., Brilliant Violet dyes), then the use of Super Bright Staining Buffer (Cat. No. SB-4400) is recommended to minimize any non-specific polymer interactions between these fluorochromes. For specific instructions for use, please refer to the product technical data sheet.

Can the Super Bright dyes be used for non-flow applications?

Super Bright dyes have not been tested for applications other than flow cytometry.

What is the difference between the Super Bright and eVolve dyes?

Super Bright dyes are fluorochromes based on polymer technology and are excited by the violet laser. In contrast, eVolve dyes are based on Qdot nanocrystal technology and are maximally excited by the UV laser, although they may also be excited by violet, blue, yellow-green, and, depending on the eVolve, the red laser lines. The general shape of the emission spectra for Super Bright dyes will be more similar to traditional fluorochromes, while eVolve dyes have extremely narrow emission spectra. Please contact Tech Support (techsupport@thermofisher.com) for more information.

What are the Super Bright dyes?

eBioscience Super Bright dyes are a series of patent-pending fluorochromes that are based upon a fluorescent polymer and its tandems. Super Bright dyes have been developed for use in flow cytometry and can be excited by the violet laser (405 nm). The peak emission of each Super Bright dye is indicated by the number in the product description, e.g., "Super Bright 600" has a peak emission of 600 nm.

Are the Super Bright Dyes photo-labile?

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.


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