DyLight™ Multiplex 649/549 Duo Western Blot Kit

Catalog No. ABIN6719664

Product Details

Reactivity: Mouse, Rabbit

Host: Goat

Application: BioImaging (BI), Immunofluorescence (IF), Immunostaining (ISt), Multiplex Assay (MA), Standard (STD), Western Blotting (WB)

Purpose: This kit is suitable for fluorescent western blotting, multiplex analysis, including multicolor imaging, utilizing various commercial gel imaging systems.

Characteristics:

Synonyms: Fluorescent western blotting, Multiplex western blotting, multi-color western blotting, fluorescent labelled antibodies, fluorescent imaging, DyLights, dyLight conjugates

Background: The The DyLight™ multiplex 649/549 Duo fluorescent Western blotting kit is suited for simultaneous detection and quantification of specific protein populations in a biological sample. Using a combination of two antibodies selected for minimal cross reactivity, fluorescent detection method enables simultaneous quantitative analysis of multiple proteins within the same sample on the same blot. The DyLight™ multiplex 649/549 Duo Western blot kit contain all the necessary components that are optimized for the simultaneous detection of multiple proteins on the same blot using DyLight™-dye labeled secondary antibodies that are visualized in different fluorescence channels (649/549). The kit also includes blocking buffer, wash buffer, pre-stained protein standard and an incubation box for convinience and ultimate performance with minimal or no optimization. The fluorescent dyes such as DyLights™ when conjugated to secondary antibodies, offer a variety of benefits over traditional detection methods such as colorimetric and chemiluminescent detection. Multiplex detection using the correct lighting and filter conditions, enables the quantitation of multiple proteins and eliminates the need to strip and reprobe. Other benefits of fluorescent Western blotting include increased sensitivity, excellent signal stability over time as well as precise quantitative analysis with broader dynamic range and high linearity. Due to their exceptional photostability, DyLight™ dye conjugates can be archived and visualized several times without a decrease in signal.

Components: DyLight™ Multiplex 649/549 Duo Western Blot Kit contains Goat anti-mouse IgG (H+L) DyLight™ 549, Goat anti-rabbit IgG (H+L) DyLight™ 649, opal pre-stained protein standard, wash buffer (10x TTBS), blocking buffer (2x) and incubation box.

Application Details

Protocol:

Production: The DyLight conjugated secondary antibodies were prepared from monospecific antiserum by immunoaffinity chromatography using Rabbit or Mouse IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Goat Serum, Rabbit or Mouse IgG and Rabbit or Mouse Serum. No reaction was observed against Bovine, Chicken, Goat, Guinea Pig, Hamster, Horse, Human, Rat and Sheep Serum Proteins. These antibodies will react with heavy chains of rabbit or mouse IgG and with light chains of most rabbit or mouse immunoglobulins. Blocking buffer is specifically formulated to achieve superior reproducible western blotting images using this system. Wash buffer (10X TTBS) was aseptically filtered through a Millipore 0.22 micron filter into clean, pre-sterilized containers. The product was tested on trypticase soy agar for 24 hours, 48 hours and 72 hours and was found to be negative for bacteria.

Restrictions: For Research Use only

Handling

Storage: RT,4 °C,-20 °C

Storage Comment: The secondary antibodies should be stored at 4°C prior to reconstitution. For extended storage aliquot contents and freeze at -20°C. Avoid freeze/thaw cycles. The wash buffer and blocking buffer can be stored at 2-8°C prior to opening. The pre-stained protein Western standards should be stored at 2-8°C. The pre-stained protein standards can also be stored at room temperature for up to 6 months or at -20°C for up to two years.

Expiry Date: 12 months