F(ab')2-Goat anti-Mouse IgG (H+L) Secondary Antibody, NovaFluor™ Yellow 660
The goat anti-mouse IgG polyclonal antibody reacts with the heavy and light chains of mouse IgG, with minimal cross reactivity to human, rat, bovine, goat, hamster, and rabbit serum proteins. Reactivity has been shown to IgG1, IgG2a, IgG2b, and IgG3, as well as some reactivity to IgM. The antibody is a F(ab')2 fragment which contains the 2 Fab domains still linked through the hinge region of the immunoglobulin. Removal of the Fc region helps to reduce issues typically found with Fc-mediated binding.IThis polyclonal has been validated as a secondary reagent (its ability to recognize cells stained with a mouse IgG followed by this polyclonal antibody).
Applications Reported: This polyclonal antibody has been reported for use in flow cytometric analysis.
Our internal testing shows that NovaFluor Yellow 660 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.
Applications Tested: This polyclonal antibody has been tested by flow cytometric analysis of normal human peripheral blood cells stained first with a mouse anti-human antibody. This can be used at less than or equal to 0.5 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. This polyclonal antibody has been tested by immunohistochemistry on formalin-fixed paraffin embedded sections that have been stained first with a mouse anti-human antibody. This can be used at less than or equal to 10 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Each NovaFluor conjugate or kit is shipped with CellBlox Blocking Buffer. Use this buffer whenever staining with NovaFluor conjugates, including single-color compensation controls using cells. Whenever possible, we recommend adding CellBlox 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 100µL of cell sample containing 10^3 to 10^8 cells.
Excitation: 552 nm; Emission: 663 nm; Laser: 561 nm (Yellow) Laser
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