Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Flow cytometry [2]
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- Product number
- H002T02U01-A - Provider product page

- Provider
- Invitrogen Antibodies
- Product name
- CD3 Monoclonal Antibody (UCHT1), NovaFluor™ UV 765, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description<br>The UCHT1 monoclonal antibody reacts with human CD3e, a 20 kDa subunit of the TCR complex. Along with the other CD3 subunits gamma and delta, the epsilon chain is required for proper assembly, trafficking and surface expression of the TCR complex. CD3 is expressed by thymocytes in a developmentally regulated manner and by all mature T cells. Crosslinking of TCR via immobilized UCHT1 initiates an intracellular biochemical pathway resulting in cellular activation and proliferation. <br><br>This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.<br><br>Applications Tested<br>This UCHT1 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.6 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.<br><br>Master mixes<br>• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.<br>• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.<br><br>Whole Blood compatibility<br>• 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.<br>• See the Bulk Lysis of Human Whole Blood protocol <a href="https://www.thermofisher.com/us/en/home/references/protocols/cell-and-tissue-analysis/protocols/red-blood-cell-lysis.html">here</a>.<br>• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.<br><br>Viability dye compatibility<br>• NovaFluor dyes are not compatible with DNA intercalating viability dyes.<br>• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. <a href="https://www.thermofisher.com/us/en/home/life-science/cell-analysis/flow-cytometry/flow-cytometry-assays-reagents/cell-viability-assays-flow-cytometry/fixable-viability-dyes-flow-cytometry.html">Invitrogen LIVE/DEAD Fixable Dead Cell stains</a> are recommended for use with NovaFluor dyes.<br><br>CellBlox Plus Blocking Buffer<br>• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (Cat. No. C001T03F01), essential for optimal staining.<br>• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.<br>• Add 5 µL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.<br>• 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.<br>• For single-color controls, use 5 µL of CellBlox Plus Blocking Buffer per 100 µL of cell sample (10^3 to 10^8 cells).<br><br>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. <a href="https://www.thermofisher.com/us/en/home/life-science/cell-analysis/flow-cytometry/antibodies-for-flow-cytometry/novafluor-dyes.html">Learn more</a><br><br>Excitation: 358 nm; Emission: 763 nm; Laser: 349 nm (Ultraviolet) Laser
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- UCHT1
- Vial size
- 25 Tests
- Concentration
- 0.6 μg/Test
- Storage
- 4°C, store in dark, DO NOT FREEZE!
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image

- Experimental details
- Spectral signature for NovaFluor UV 765 collected on a 5-laser Cytek Aurora Full Spectrum flow cytometer using Cytek assay settings. Human peripheral blood mononuclear cells were stained with anti-human CD4 (SK3) and signatures displayed following gating on the lymphocyte population.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image

- Experimental details
- Normal human peripheral blood cells were unstained (left) or stained with CD3 Monoclonal Antibody, NovaFluor UV 765 (right). All cells were co-stained with CD19 Monoclonal Antibody, eFluor 450 (Product # 48-0199-42). 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 multiple autofluorescence signature extraction.