Invitrogen™ CD10 Monoclonal Antibody (eBioCB-CALLA (CB-CALLA)), Super BrighT™ 600, eBioscience™

Catalog No.	Quantity
63010642	100 Tests

Catalog No. 63010642 Supplier Invitrogen™ Supplier No. 63010642

Description

Mouse Monoclonal Antibody

Description: The eBioCB-CALLA monoclonal antibody recognizes human CD10 (CALLA, NEP, enkephalinase, Neprilysin), which is a 100 kDa, type II cell surface glycoprotein originally identified for its expression on most acute lymphoblastic leukemias (ALL). Subsequently, CD10 was shown to be the same molecule as the neutral endopeptidase (NEP), or KII-NA. CD10 is a Zn2+-dependent metallo-peptidase with endothelin, glucagon, gastrin, neurotensin and bradykinin included among its substrates. CD10 is involved in the regulation of chemotactic and inflammatory processes involving neutrophils. In B cells, CD10 regulates stromal cell-dependent B lymphopoiesis and expression has also been reported on mature B cells in germinal centres. In addition to the hematopoietic compartment, other major sites of CD10 expression are the brush border of enterocytes and renal tubules and glomeruli. There is partial blocking of the eBioCB-CALLA and MEM-78 monoclonal antibodies indicating that they recognize similar epitopes. Applications Reported: This CB-CALLA antibody has been reported for use in flow cytometric analysis. Applications Tested: This CB-CALLA antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 μL (0.06 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...

CD10, also known as the Common Acute Lymphocytic Leukemia Antigen (CALLA) and neutral endopeptidase (NEP), is a Zn2+-dependent metallo-peptidase with neutral metalloendopeptidase activity. It is a 100 kDa type II transmembrane glycoprotein encoded by a gene that exists in a single copy of greater than 45 kb. The 5' untranslated region of the CD10 gene is alternatively spliced, resulting in four separate mRNA transcripts, although the coding region remains unaffected by alternative splicing. CD10 is involved in the regulation of chemotactic and inflammatory processes involving neutrophils and plays a role in stromal cell-dependent B lymphopoiesis. It is expressed on immature B cells in adult bone marrow, mature B cells in germinal centers, and cells from patients with chronic myelocytic leukemia (CML). Additionally, CD10 is present on the cells of lymphoblastic, Burkitt's, and follicular germinal center lymphomas. Beyond the hematopoietic compartment, CD10 is highly expressed on the brush border of enterocytes and renal tubules and glomeruli, as well as on breast myoepithelial cells, bile canaliculi, and fibroblasts. CD10 cleaves peptides at the amino side of hydrophobic residues, inactivating several biologically active peptides, including endothelin, glucagon, gastrin, neurotensin, bradykinin, enkephalins, substance P, oxytocin, and others. Diseases associated with CD10 dysfunction include spinocerebellar ataxia 43 and Charcot-Marie Tooth Disease, highlighting its significance in both normal physiological processes and disease states.

Specifications

• Antigen: CD10
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: eBioCB-CALLA (CB-CALLA)
• Concentration: 5 μL/Test
• Conjugate: Super Bright 600
• Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
• Gene: Mme
• Gene Accession No.: P08473
• Gene Alias: 6030454K05Rik; Atriopeptidase; C85356; CALLA; CD10; CMT2T; common acute lymphoblastic leukemia antigen; common acute lymphocytic leukemia antigen; Enkephalinase; EPN; membrane metallo endopeptidase; membrane metalloendopeptidase; membrane metallo-endopeptidase; membrane metallo-endopeptidase (neutral endopeptidase, enkephalinase, CALLA, CD10); membrane metallo-endopeptidase (neutral endopeptidase/enkephalinase); membrane metallo-endopeptidase variant 1; membrane metallo-endopeptidase variant 2; MME; NEP; neprilysin; neprilysin-390; neprilysin-411; neutral endopeptidase 24.11; SFE; Skin fibroblast elastase
• Gene Symbols: Mme
• Host Species: Mouse
• Purification Method: Affinity chromatography
• Quantity: 100 Tests
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 4311
• Target Species: Human
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG2b 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.