Usage
Neutralization of Bioactivity, Flow Cytometry, ELISA capture. Neutralization of Human CCL13 Bioactivity: The exact concentration of antibody required to neutralize human CCL13 activity is dependent on the cytokine concentration, cell type, growth conditions and the type of activity studied. The Neutralization Dose50 (ND50) for this antibody is defined as that concentration of antibody required to yield one-half maximal inhibition of the cytokine activity on a responsive cell line, when that cytokine is present at a concentration just high enough to elicit a maximum response. Flow Cytometry: This antibody was validated for flow cytometry using IL-1 treated A549 cells. For intracellular staining to detect human CCL13, cells must first be fixed and permeabilized using 4% paraformaldehyde and 0.1% saponin in PBS. Dilute this antibody to 25 ug/mL and add 10 ul of the diluted solution to 1-2.5 x 10^5 cells in a total reaction volume not exceeding 200ul. The binding of unlabeled monoclonal antibodies may be visualized by adding 10 ul of a 25 ug/mL stock solution of a secondary developing reagent such as goat anti-mouse IgG conjugated to a fluorochrome. ELISA Capture: This antibody can be used as a capture antibody in a human CCL13 ELISA in combination with the biotinylated polyclonal detection antibody. A general protocol is provided on the next page. Using plates coated with 100 ul/well of the capture antibody at 2 ug/mL, in combination with 100 ul/well of the detection antibody, an ELISA for sample volumes of 100 ul can be obtained. To arrive at the optimal dose range for this ELISA, set up a two-fold dilution series of the protein standard starting with 0.5 ng/ml.
Presentation
Lyophilized from PBS, 5% Trehalose
Reconstitution
Phosphate Buffered Saline (PBS)
Restrictions
For research use only. Intended for use by laboratory professionals.
Guarantee
This antibody carries the LSBio 100% Guarantee.
LSBio Guarantee