Invitrogen™ CD150 Monoclonal Antibody (mShad150), Super Bright™ 645, eBioscience™, Invitrogen™

Catalog No.	Quantity
64-150-282	100 μg

Catalog No. 64-150-282 Supplier Invitrogen™ Supplier No. 64150282

Description

Rat Monoclonal Antibody

Description: The eBioGHI/61 monoclonal antibody reacts with human CD163 (Haemoglobin Scavenger Receptor, M130), a member of the macrophage scavenger receptor family. CD163 is expressed by tissue macrophages, circulating blood monocytes, and some reports suggest that it may also be expressed by haematopoietic progenitor cells. CD163 functions as a high affinity scavenger receptor for the complex of haemoglobin and haptoglobin which forms to clear haemoglobin from the blood. The high affinity for CD163 is specific for the haemoglobin-haptoglobin complex, whereas haemoglobin or haptoglobin display no to low affinity toward CD163. Upon ligand binding, CD163 initiates signal transduction which leads to the production of IL-6 and IL-10. Furthermore, it has been demonstrated that cytokines such as IL-6 and IL-10 can induce expression of CD163, whereas proinflammatory stimuli, such as LPS or TNF alpha, can cause shedding of a soluble form of CD163, whose function is unclear. Applications Reported: This eBioGHI/61 (GHI/61) antibody has been reported for use in flow cytometric analysis. Applications Tested: This eBioGHI/61 (GHI/61) antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 μL (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.

CD150, also known as SLAM (Signaling Lymphocyte Activation Molecule), is a member of the immunoglobulin gene superfamily and plays a crucial role in T cell stimulation and immune cell interactions. It is constitutively expressed on peripheral blood memory T cells, T cell clones, immature thymocytes, and a proportion of B cells. CD150 expression is rapidly upregulated upon activation, particularly on naive T cells, Th1 cells, and activated B cells, which express the membrane-bound, soluble, and cytoplasmic isoforms of SLAM. In T cells, signaling through SLAM induces proliferation and enhances the interferon-gamma response. SLAM is also involved in adhesion between T cells and antigen-presenting cells, facilitating immune cell interactions. Homophilic SLAM-SLAM binding during B to B and B to T cell interactions is suggested to enhance the expansion and differentiation of activated B cells. CD150 serves as a receptor for the measles virus and acts as a co-activator on T and B cells. It interacts with SH2D1A and PTPN11 via its cytoplasmic domain, playing a role in signal transduction. Mutations in the CD150 gene may be associated with X-linked lymphoproliferative disease (XLP), highlighting its importance in immune regulation and disease pathology.

Specifications

• Antigen: CD150
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: mShad150
• Concentration: 0.2 mg/mL
• Conjugate: Super Bright 645
• Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
• Gene: Slamf1
• Gene Accession No.: Q9QUM4
• Gene Alias: 4933415F16; AA177906; CD150; CD150 antigen; CDw150; ESTM51; IPO-3; RGD1560634; signaling lymphocyte activation molecule; signaling lymphocytic activation molecule; signaling lymphocytic activation molecule family member 1; Slam; SLAM family member 1; Slamf1
• Gene Symbols: Slamf1
• Host Species: Rat
• Purification Method: Affinity chromatography
• Quantity: 100 μg
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 27218
• Target Species: Mouse
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG2b λ

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.