Invitrogen™ HLA-DR Monoclonal Antibody (LN3), Super Bright™ 780, eBioscience™

Catalog No.	Quantity
78-995-642	100 Tests

Catalog No. 78-995-642 Supplier Invitrogen™ Supplier No. 78995642

Description

Mouse Monoclonal Antibody

Description: The LN3 mAb reacts with the human major histocompatibility complex (MHC) class II, HLA-DR. HLA-DR is expressed on the surface of human antigen presenting cells (APC) including B cells, monocytes, macrophages, DCs, and activated T cells. HLA-DR is a heterodimeric transmembrane protein composed of alpha and beta subunits and plays an important role in the presentation of peptides to CD4+ T lymphocytes. Applications Reported: This LN3 antibody has been reported for use in flow cytometric analysis. Applications Tested: This LN3 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 μL (0.125 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 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter. 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 Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information. In some experiments, we have observed that compensat...

HLA-DR, like other MHC class II molecules, is a transmembrane glycoprotein composed of a 36 kDa alpha chain (DRA) and 27 kDa beta chain (DRB). The alpha chain gene contains 5 exons. Exon 1 encodes the leader peptide, exons 2 and 3 encode the two extracellular domains, and exon 4 encodes the transmembrane domain and the cytoplasmic tail. DRA does not have polymorphisms in the peptide binding part and acts as the sole alpha chain for DRB1, DRB3, DRB4 and DRB5. Within the DR molecule the beta chain contains all the polymorphisms specifying the peptide binding specificities. Hundreds of DRB1 alleles have been described and typing for these polymorphisms is routinely done for bone marrow and kidney transplantation. HLA-DR is expressed primarily on antigen presenting cells such as B lymphocytes, monocytes, macrophages, thymic epithelial cells and activated T lymphocytes. Three loci, DR, DQ and DP, encode the major expressed products of the human class II region. The human MHC class II molecules bind intracellularly processed peptides, present them to T-helper cells, and have a critical role in the initiation of the immune response. 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-DR
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: LN3
• Concentration: 5 μL/Test
• Conjugate: Super Bright 780
• Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
• Gene: CD74
• Gene Accession No.: P01903, P01911, P04233, P13762, P79483, Q30154
• Gene Alias: AI323765; BLA-DRB3; BOLA-DRA; BoLA-DR-alpha; BoLA-DRB; BOLA-DRB3; BoLA-DRB3 protein; BoLA-DRB3.2; Bota-DRB01; Bota-DRB02; Bota-DRB04; Bota-DRB07; Bota-DRB21; Bota-DRB22; bovine leukocyte antigen; Cd74; CD74 antigen; CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated); CD74 molecule; Cd74 molecule, major histocompatibility complex, class II invariant chain; class II-associated invariant chain peptide; CLIP; Clone P2-beta-3; DASS-397D15.1; DHLAG; dinucleotide microsatellite; DLA class II histocompatibility antigen, DR-1 beta chain; DLA DRBB1 beta chain; DLA-DR beta; DLA-DRB1; DLA-DRBB1; DR beta-5; DR beta-chain antigen binding domain; DR-16; DR2-beta-2; DR4; DR-4; DR7; DR9; DR-9; DRB; DRB1; DRB1 transplantation antigen; DRB3; DRB4; DRBB1; DR-beta chain; DR-beta chain MHC class II; DRw10; Dw2; DW2.2/DR2.2; E-alpha-f; FLJ51114; FLJ75017; FLJ76359; gamma chain of class II antigens; H-2 class II histocompatibility antigen gamma chain; H-2 class II histocompatibility antigen, E-D alpha chain; H-2 class II histocompatibility antigen, E-K alpha chain; H-2 class II histocompatibility antigen, E-U alpha chain; H-2Ea; H2-Ea; H2-Ea-ps; H2-IE-alpha; histocompatibility 2, class II antigen E alpha; histocompatibility 2, class II antigen E alpha, pseudogene; histocompatibility antigen HLA-DR alpha; histocompatibility complex, class II, DR beta 3; histocompatibility: class II antigens, gamma chain of; HLA class II histocompatibility antigen gamma chain; HLA class II histocompatibility antigen, DR alpha chain; HLA class II histocompatibility antigen, DR beta 3 chain; HLA class II histocompatibility antigen, DR beta 4 chain; HLA class II histocompatibility antigen, DR beta 5 chain; HLA class II histocompatibility antigen, DR-1 beta chain; HLA class II histocompatibility antigen, DR-5 beta chain; HLA class II histocompatibility antigen, DRB1-15 beta chain; HLA class II histocompatibility antigen, DRB1-16 beta chain; HLA class II histocompatibility antigen, DRB1-3 chain; HLA class II histocompatibility antigen, DRB1-7 beta chain; HLA class II histocompatibility antigen, DRB1-9 beta chain; HLADG; HLA-DR antigens-associated invariant chain; HLA-DR1B; HLA-DR3B; HLA-DR4B; HLA-DRA; HLA-DRA1; HLA-DRB; HLA-DRB1; HLA-DRB2; HLA-DRB3; HLA-DRB4; HLA-DRB5; HLA-DR-gamma; human leucocyte antigen DRB1; human leucocyte antigen DRB3; human leucocyte antigen DRB4; human leucocyte antigen DRB5; ia antigen-associated invariant chain; Ia3; Ia-3; Ia-associated invariant chain; Ia-GAMMA; I-E alpha MHC class II; I-Ealpha; II; integral membrane glycoprotein; invariant gamma chain; invariant polypeptide of major histocompatibility complex, class II antigen-associated; INVG34; LA-DRB; leukocyte antigen; leukocyte antigen class II; leukocyte antigen DRB3; lymphocyte antigen DRB1; major histocompatibility complex class II DR-beta chain; major histocompatibility complex, class II, DR alpha; major histocompatibility complex, class II, DR alpha precursor; major histocompatibility complex, class II, DR beta 1; major histocompatibility complex, class II, DR beta 3; major histocompatibility complex, class II, DR beta 4; major histocompatibility complex, class II, DR beta 4 precursor; major histocompatibility complex, class II, DR beta 5; major histocompatibility complex, class II, DRB3; MHC cell surface glycoprotein; MHC class I antigen; MHC class II antigen; MHC class II antigen beta chain; MHC class II antigen BoLA-DRB3; MHC class II antigen DR beta 3 chain; MHC class II antigen DRA; MHC class II antigen DRB1*15; MHC class II antigen DRB1*16; MHC class II antigen DRB1*3; MHC class II antigen DRB1*9; MHC class II antigen DRB3; MHC class II antigen DRB4; MHC class II antigen DRB5; MHC class II antigen E alpha; MHC class II antigen HLA-DR-beta; MHC class II DLA DRB1 beta chain; MHC class II DLA-DRB; MHC class II DLA-DR-beta-1; MHC class II DR beta 1; MHC class II DR beta chain; MHC class II DR beta-chain
• Gene Symbols: CD74, HLA-DRA, HLA-DRB1, HLA-DRB3, HLA-DRB4, HLA-DRB5
• Host Species: Mouse
• Purification Method: Affinity chromatography
• Quantity: 100 Tests
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 3122, 3123, 3125, 3126, 3127, 972
• 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.

Safety and Handling

WARNING: Cancer - www.P65Warnings.ca.gov