Rabbit IgG Isotype Control [Alexa Fluor® 647] Summary
This Rabbit IgG immunoglobulin is useful as an isotype control for Rabbit IgG antibody.
Rabbit IgG whole molecular was prepared from normal serum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer. Rabbit IgG whole molecular was assayed by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit IgG and anti-Rabbit Serum.
Read 1 Review rated 3 using NBP2-36463AF647 in the following application:
1:100 dilution in 1X Permeabilization buffer (Foxp3 Staining Buffer Set from eBiosciences), 30 minutes incubation at 21°C
LSR II flow cytometer (BD); NBP1-49533AF647 stained Mouse CD4+ T Cells as +ve Control and Fluorescence minus one/FMO control
See Additional Comments
Cells fixed and permeabilized with Foxp3 Staining Buffer set (eBioSciences #00-5523-00) for intracellular staining
One wash with 1X Permeabilization buffer (eBiosciences) and one wash with FACS buffer (1x PBS, 1% BSA, 0.1% sodium azide)
Murine spleens were harvested and CD4+ T cells were isolated via negative selection. CD4+ T cells were stimulated with anti-CD3/CD28 beads and insulin (1μg/mL) for 5 days in culture media with additional glucose provided. Cells were washed and beads removed prior to staining. Cells were stained for surface antigens, washed, then fixed and permeabilized (via Foxp3 Staining Buffer set (eBioSciences #00-5523-00)) for intracellular staining (Novus informed me that the epitope is intracellular for this antibody). Cells were washed again before being ran on LSR II Flow Cytometer. Data was analyzed via FlowJo (Tree Star). Cells were gated, in order, on lymphocytes, single cells, live cells, and CD4+ cells. Positive controls and both isotype and FMO controls were compared.
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Understanding the Importance of Human IgG Human IgG is a component of the immune system that protects the body from infection. It is the most abundantly found antibody isotype within the circulatory system of the human body. All antibody isotypes contain two heavy chains and two light chains... Read full blog post.
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