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Invitrogen
MHC Class I (H-2Db) Monoclonal Antibody (28-14-8), NovaFluor™ Yellow 730, eBioscience™
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MHC Class I (H-2Db) Antibody (M038T03Y07-A) in Flow
Product Details
M038T03Y07-A
Species Reactivity
Host/Isotype
Class
Type
Clone
Conjugate
Excitation/Emission Max
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Concentration
Purification
Storage buffer
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RRID
Product Specific Information
Description
The 28-14-8 monoclonal antibody reacts with the mouse MHC class I, H-2D^b, and cross-reacts with H-2L^d, H-2D^q and/or H-2L^q. Binding of 28-14-8 to the alpha 3 domain of H-2L^d is not dependent on beta2-microglobulin.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
The 28-14-8 antibody has been tested by flow cytometric analysis of mouse splenocytes and can be used at less than or equal to 1 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.
Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.
Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.
Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.
CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 µL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 µL of CellBlox Plus Blocking Buffer per 100 µL of cell sample (10^3 to 10^8 cells).
NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more
Our internal testing shows that NovaFluor Yellow 730 non-specifically stains B cells in SJL mice. Non-specific staining has not been observed in BALB/c or C57BL/6 mice. Other strains have not been tested. See the Antibody Testing Data for an example of this strain-dependent difference.
Excitation: 551 nm; Emission: 730 nm; Laser: 561 nm (Yellow) Laser
Target Information
This protein is an MHC class I alloantigen, which is involved in antigen presentation to T cells expressing CD3/TCR and CD8.
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
How to use the Panel Builder
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
References (0)
Bioinformatics
Protein Aliases: H-2 class I histocompatibility antigen, D-B alpha chain; H-2D cell surface glycoprotein; H-2D(B); H2-DC1-beta transplantation antigen protein; H2D1; H2DB; H2L; H2LD; MHC class I antigen; MHC class I H-2Dp; MHC class I H2 antigen; MHC H2-D-q alpha-chain; Q5k protein
Gene Aliases: H-2D; H2-D; H2-D1
UniProt ID: (Mouse) P01899
Entrez Gene ID: (Mouse) 14964
Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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