H019T03B12-A

antibody from Invitrogen Antibodies

Targeting: CD14

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

H019T03B12-A

Provider

Invitrogen Antibodies

Product name

CD14 Monoclonal Antibody (61D3), NovaFluor™ Blue 800, eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The 61D3 monoclonal antibody reacts with human CD14, a 53-55 kDa GPI-linked glycoprotein. CD14 is expressed on monocytes, interfollicular macrophages and some dendritic cells. Complexes of LPS and LBP (LPS-Binding Protein) bind with high affinity to monocytes through the surface CD14. Applications Reported: The 61D3 antibody has been reported for use in flow cytometric analysis. Applications Tested: The 61D3 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.4 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. Each product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer . 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. This NovaFluor conjugate has been updated to ship with CellBlox Plus Blocking Buffer (Cat. No. (C001T06F01)). This buffer contains formulation improvements over CellBlox. CellBlox Plus Blocking Buffer is required for optimal staining with NovaFluor conjugates and should be used in all experiments where NovaFluor conjugates are used. Whenever possible, we recommend adding CellBlox Plus Blocking Buffer to antibody cocktails/master mixes prior to combining with cells. Add 5 µL per sample (regardless of the number of NovaFluors in your panel) to use the antibody cocktail as intended. For single-color controls, use 5 µL of CellBlox Blocking Buffer per 100 µL of cell sample containing 10^3 to 10^8 cells. NovaFluor conjugates are based on Phiton technology utilizing novel nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more Excitation: 493nm; Emission: 806 nm; Laser: 488 nm (Blue) Laser

Reactivity

Human

Host

Mouse

Conjugate

Infrared dye

Isotype

IgG

Antibody clone number

61D3

Vial size

100 Tests

Concentration

0.4 µg/Test

Storage

4°C, store in dark, DO NOT FREEZE!

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Spectral signature for NovaFluor Blue 800 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.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Normal human peripheral blood cells were either left unstained (blue histogram) or stained with CD14 Monoclonal Antibody, NovaFluor Blue 800 (purple histogram). Total viable cells in the monocyte 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.