Invitrogen™ HLA-ABC Monoclonal Antibody (W6/32), Super Bright™ 436, eBioscience™, Invitrogen™

Catalog No.	Quantity
62-998-342	100 Tests

Catalog No. 62-998-342 Supplier Invitrogen™ Supplier No. 62998342

Description

Mouse Monoclonal Antibody

Description: The W6/32 monoclonal antibody reacts with the human major histocompatibility complex (MHC) class I, HLA-A, B, C. MHC class I antigens associated with beta 2-microglobulin are expressed by all human nucleated cells and are central in cell-mediated immune response and tumor surveillance. W6/32 mAb recognizes a non-polymorphic epitope shared among products of the HLA-A, B, and C loci and immunoprecipitates both 43 kDa and 11-12 kDa chains. Crossreactivity is also seen in baboon, rhesus and cynomolgus monkey. Applications Reported: This W6/32 antibody has been reported for use in flow cytometric analysis. Applications Tested: This W6/32 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 μL (0.5 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. Super Bright 436 can be excited with the violet laser line (405 nm) and emits at 436 nm. We recommend using a 450/50 bandpass filter, or equivalent. Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Staining Buffer (Product No. SB-4400) to minimize any non-specific polymer interactions.

HLA-class I major histocompatibility (MHC) antigens are intrinsic membrane glycoproteins expressed on nucleated cells and noncovalently associated with an invariant beta2 microglobulin. They carry foreign determinants important for immune recognition by cytotoxic T cells, thus important for anti-viral and anti-tumor defense. Human HLA-class I antigens are represented by HLA-A, HLA-B and HLA-C molecules. HLA and MHC antibodies play a significant role in Immunopeptidomics, facilitating the identification and characterization of neoantigens through high-performance liquid chromatography coupled to tandem Mass Spectrometry.

Specifications

• Antigen: HLA-ABC
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: W6/32
• Concentration: 5 μL/Test
• Conjugate: Super Bright 436
• Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
• Gene: HLA-B
• Gene Accession No.: P01889, P04439, P10321
• Gene Alias: A29 protein; alpha chain of MHC class II; antigen presenting molecule; AS; Aw-33; Aw-74; B-4901; B-5001; Bw-47; Bw-50; class I MHC, HLA-B antigen; D6S204; epididymis secretory protein Li 83; H-2K; H2-K; H-2K(d); HEL-S-83; histocompatibility antigen; HLA A; HLA -B; HLA B-40011; HLA B44; HLA C; HLA class I antigen; HLA class I antigen HLA-B; HLA class I histocompatibility antigen C alpha chain; HLA class I histocompatibility antigen Cw-1 alpha chain; HLA class I histocompatibility antigen, A alpha chain; HLA class I histocompatibility antigen, A-1 alpha chain; HLA class I histocompatibility antigen, A-11 alpha chain-like; HLA class I histocompatibility antigen, A-30 alpha chain; HLA class I histocompatibility antigen, A-33 alpha chain; HLA class I histocompatibility antigen, A-74 alpha chain; HLA class I histocompatibility antigen, B alpha chain; HLA class I histocompatibility antigen, B-47 alpha chain; HLA class I histocompatibility antigen, B-50 alpha chain; HLA class I histocompatibility antigen, B-82 alpha chain; HLA class I histocompatibility antigen, C alpha chain; HLA class I histocompatibility antigen, Cw-1 alpha chain; HLA class I histocompatibility antigen, Cw-15 alpha chain; HLA class I histocompatibility antigen, Cw-18 alpha chain; HLA locus; HLA molecule; HLAA; HLA-A; HLA-A locus alpha 2 domain; HLA-A null; HLA-A11; HLA-A33; HLA-Aw33.1; HLAB; HLA-B; HLA-B 1502; HLA-B alpha chain; HLA-B alpha chain (B*2706); HLA-B alpha chain (B*5002); HLA-B alpha-chain; HLA-B*45ZJ; HLA-B15; HLA-B35; HLA-B35 antigen; HLA-B-3506; HLA-B39; HLA-B-3905; HLA-B49; HLA-B50; HLA-B55; HLA-B-5502; HLA-B-5602; HLA-B59; HLA-B61; HLA-Bw50 antigen; HLAC; HLA-C; HLA-C alpha chain; HLA-C alpha chain (HLA-Cw*15v); HLA-C alpha chain for Cw*18GB; HLA-C alpha-chain; HLA-C antigen; HLA-Cw; HLA-Cw*04GB; HLA-Cw*1702; HLA-Cw*18GB; HLA-Cw-1503; HLA-DQB1; HLA-DRB1; HLA-JY3; HLC-C; Human leukocyte antigen A; Human leukocyte antigen C; human leukocyte antigen-C alpha chain; hypothetical protein; lecocyte antigen B; leucocyte antigen A; Leukocyte Antigen; leukocyte antigen C; leukocyte antigen class I-A; leukocyte antigen class I-B; Leukoocyte Antigen; LOC100515902; LOW QUALITY PROTEIN: HLA class I histocompatibility antigen, A-11 alpha chain-like; lymphocyte antigen; major histocompatability complex B antigen; major histocompatibility antigen HLA-C; major histocompatibility complex, class I, A; major histocompatibility complex, class I, B; major histocompatibility complex, class I, C; MCH class I antigen; MGC184092; MGC7052; MHC; MHC 1; MHC antigen; MHC clas I antigen; MHC Class 1; MHC class 1 antigen; MHC class I; MHC class I alpha chain; MHC class I anitgen; MHC class I anti; MHC Class I antigen; MHC class I antigen A*30; MHC class I antigen A*33; MHC class I antigen A*74; MHC class I antigen B*47; MHC class I antigen B*50; MHC class I antigen B*82; MHC class I antigen Cw*15; MHC class I antigen Cw*18; MHC class I antigen GN00104; MHC class I antigen heavy chain; MHC class I antigen heavy chain HLA-C; MHC class I antigen HLA-A heavy chain; MHC class I antigen HLA-A33; MHC class I antigen HLA-B alpha chain; MHC class I antigen HLA-B heavy chain; MHC class I antigen null protein; MHC class I antigen SHCHA; MHC class I antigene; MHC class I histocompatibility antigen; MHC class I human leukocyte antigen; MHC class I molecule; MHC class I protein; MHC class II antigen; MHC class l antigen; MHC classI antigen; MHC HLA-ABC; MHC HLA-B cell surface glycoprotein; MHC HLA-B transmembrane glycoprotein; MHC HLA-B27d; MHC HLA-B27-HS; MHC HLA-B39N; MHC I HLA; MHC1; PSORS1; SPDA1; surface antigen; truncated class I antigen; truncated MHC class I antigen
• Gene Symbols: HLA-A, HLA-B, HLA-C
• Host Species: Mouse
• Purification Method: Affinity chromatography
• Quantity: 100 Tests
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 3105, 3106, 3107
• Target Species: Human
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG2a 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.