Invitrogen™ CD127 Monoclonal Antibody (eBioRDR5), Super Bright™ 702, eBioscience™, Invitrogen™

Catalog No.	Quantity
67-127-842	100 Tests

Catalog No. 67-127-842 Supplier Invitrogen™ Supplier No. 67127842

Description

Mouse Monoclonal Antibody

Description: The 13A4 monoclonal antibody recognizes mouse Prominin-1 (sometimes also referred to as CD133 and, in the case of the human orthologue, as AC133), a 115-120 kDa pentaspan transmembrane (5-TM) domain glycoprotein. Prominin-1 is expressed on primitive cells such as hematopoietic stem and progenitor cells, neural and endothelial stem cells, retina and retinoblastoma, as well as developing epithelium. To date, the function and ligand of Prominin-1 are unknown. The 13A4 antibody does not cross react with rat, human, chicken, or Drosophila antigen but has been reported to work in canine/dog. Applications Reported: This 13A4 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 13A4 antibody has been tested by flow cytometric analysis of mouse bone marrow cells. This may be used at less than or equal to 0.5 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. Super Bright 702 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 702 nm. We recommend using a 710/50 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.

CD127, also known as the Interleukin-7 receptor alpha chain (IL-7Ralpha), is a type I glycoprotein with a molecular weight of 75-80 kDa. It forms a multi-functional receptor complex with CD132, the common gamma chain, to create the IL-7 receptor (IL-7R), which is crucial for lymphopoiesis. The active IL-7 receptor is an alpha/gamma chain heterodimer, where the gamma chain also associates with the interleukin-2 receptor and primarily activates signal transduction, while the alpha chain determines specific signaling events through its association with cytoplasmic signaling molecules. CD127 is expressed by immature B cells through the early pre-B stage, thymocytes during various stages of development, and most mature T cells, with transient down-regulation upon activation. It promotes the proliferation of precursor B cells, thymocytes, T cell progenitors, and mature CD4+ and CD8+ T cells. Binding of IL-7 to CD127 results in signal transduction through several tyrosine kinase pathways, including the Jak/STAT pathway, and is indispensable for lymphocyte development and the control of homeostatic proliferation of T cells in the periphery. Additionally, IL-7R signaling is involved in the regulation of T cell receptor (TCR) locus rearrangement in gamma delta T cells. Interestingly, CD127 expression is down-regulated on CD4+CD25+ regulatory T cells (T regs). While CD4 and CD25 co-expression is commonly used to identify T regs, this method may include cells without suppressive activity. CD4+CD25+ cells that have down-regulated CD127 expression are significantly enriched for the regulatory T cell population, as defined by the expression of the T reg-specific transcription factor Foxp3 and their suppressive activity in vitro. Diseases associated with CD127 dysfunction include severe combined immunodeficiency and T cell negative/B cell negative/NK positive severe combined immunodeficiency.

Specifications

• Antigen: CD127
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: eBioRDR5
• Concentration: 5 μL/Test
• Conjugate: Super Bright 702
• Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
• Gene: Il7r
• Gene Accession No.: P16871
• Gene Alias: CD127; CD127 antigen; CDW127; IL 7 receptor; IL 7R a; IL 7R subunit alpha; IL 7R α; IL 7Ra; IL 7Rα; IL7 receptor; IL-7 receptor alpha chain; IL7 receptor subunit alpha; IL-7 receptor subunit alpha; IL7R; IL-7R; IL7R a; IL7R alpha; IL7R subunit alpha; IL-7R subunit alpha; IL7R α; IL7RA; IL-7RA; IL-7Ralpha; IL-7R-alpha; IL7Rα; ILRA; interleukin 7 receptor; interleukin 7 receptor alpha chain; interleukin 7 receptor isoform H5-6; Interleukin7 receptor subunit alpha; interleukin-7 receptor subunit alpha; MGC107557; sCD127; soluble IL 7 receptor; soluble IL 7R
• Gene Symbols: Il7r
• Host Species: Mouse
• Purification Method: Affinity chromatography
• Quantity: 100 Tests
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 3575
• Target Species: Human
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG1 κ

Frequently Asked Questions (FAQs)

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.