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Invitrogen
MHC Class I (H-2Db) Monoclonal Antibody (28-14-8), NovaFluor™ Blue 690, eBioscience™
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FIGURE: 1 / 1
MHC Class I (H-2Db) Antibody (M038T03B09-A) in Flow
C57BL/6 mouse splenocytes were either left unstained (blue histogram) or stained with 1 .0 µg of MHC Class I (H-2Db) Monoclonal Antibody, NovaFluor Blue 690 (purple histogram). Total viable cells in the lymphocyte gate were used for analysis, as determined by LIVE/DEAD Blue (Product # L34962). Data was acquired on a 5-laser Cytek Aurora and unmixed with autofluorescence extraction.
Product Details
M038T03B09-A
Product Specifications
Species Reactivity
Mouse
Host/Isotype
Mouse
/ IgG2a, kappa
Class
Monoclonal
Type
Antibody
Clone
28-14-8
Conjugate
Excitation/Emission Max
672/688 nm
View spectra
Form
Liquid
Concentration
0.1 mg/mL
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
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 (for 25 µg or 25 test products, Cat. No. C001T02F01; for 100 µg or 100 test, 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
Excitation: 494 nm; Emission: 688 nm; Laser: 488 nm (Blue) 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.
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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
positive regulation of T cell mediated cytotoxicity
immune system process
antigen processing and presentation of peptide antigen via MHC class I
antigen processing and presentation of endogenous peptide antigen via MHC class I via ER pathway, TAP-dependent
immune response
antigen processing and presentation
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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