UniversalTM One-step Cell-based ELISA Kit (For R&D Use only)

UniversalTM One-step Cell-based ELISA Kit
Product Name & Catalog Number



















Catalog #


Product Name


Size


Content
E-R80
UniversalTM One-step Cell-bases ELISA Kit-R


With TMB Substrate


(Suitable for rabbit primary antibodies)


1 kit (1000 tests)


Fixatives (50ML)


Pretreatment Solution (50ML),


Enhancer (50ML),


50X ELISA Probe-Ro (1ML),


10X Rapid Wash Solution (100ML)


100XTMB Solution Set (2ML),


E-M80


UniversalTM One-step Cell-bases ELISA Kit-M


With TMB Substrate


(Suitable for rabbit primary antibodies)


1 kit (1000 tests)


Fixatives (50ML)


Pretreatment Solution (50ML),


Enhancer (50ML),


50X ELISA Probe-Mo (1ML),


10X Rapid Wash Solution (100ML)


100XTMB Solution Set (2ML),


E-G80


UniversalTM One-step Cell-bases ELISA Kit-G


With TMB Substrate


(Suitable for rabbit primary antibodies)


1 kit (1000 tests)


Fixatives (50ML)


Pretreatment Solution (50ML),


Enhancer (50ML),


50X ELISA Probe-Go (1ML),


10X Rapid Wash Solution (100ML)


100XTMB Solution Set (2ML),


Product Description:

UniversalTM One-step Cell-based ELISA is a latest breakthrough technology (patent pending) for directly detecting protein expression inside cells. Unlike regular (classical) ELISA, the One-step Cell-based ELISA is not only no needs of making cell lysat and coating plate but also replaces blocking, primary antibody binding, washing and secondary antibody binding with a rapid one-step reaction. Therefore, using UniversalTM One-step Cell-based ELISA kit, you can easily get your protein expression results just around one hour. The One-step Cell-based ELISA kit is a universal kit, which is suitable for most primary antibodies and no need of secondary antibody. Currently, we provide three types of UniversalTM One-step Cell-based ELISA kit: Kit-R, Kit-M and Kit-G. The Kit-R is suitable for using together with rabbit primary antibodies, the Kit-M is suitable for using together with mouse primary antibodies, and the Kit-G is suitable for using together with goat primary antibodies. Each kit is sufficient to perform 1000 assays in 96-well plates.

Application:

Direct detection of protein expression in cells.

Features:

Easy: No plate-coating, no cell-lysat-preparing, and no need of secondary antibody.

Rapid: Get results around one hour.

Universal: Suitable for most primary antibodies and no need of secondary antibody.

Reproducible: Highly reproducible results.

Storage: Enhancer: for continuous use, store in 2~8íŠ for up to 3 month, or store in -20íŠ for up to a year.

50X ELISA Probe: for continuous use, store in -20íŠ for up to 6 months or store in -70íŠ manual defrost freezer for up to a year. Repeated freezing and thawing is not recommended.

Fixatives, Pretreatment Solution and Rapid Wash Solution: store in room temperature for up to one year.

Note: Kit will be shipped in ambient temperature.

Protocol:

Materials required but not included in the kit:

Primary Antibody: Affinity-purified IgG Antibody or Monoclonal Antibody is preferred. Note: Kit-R is only suitable for rabbit primary antibodies, Kit-M is only suitable for mouse primary antibodies and Kit-G is only suitable for goat primary antibodies.

Stop solution: 1N HCl or 1N H2SO4.

Solid Phase: 96-well cell culture plate.

PBS: Phosphate Buffered Saline.

Pretreatment:

Fixing cells: If testing cells are attached cells, directly remove medium from tissue culture wells; If testing cells are suspension cells, spin down cells before removing medium). Add 50ul of Fixatives (provided in the kit) to each well (for a 96-well plate) and incubate the cells for 20 min then empty wells and rinse them twice with PBS (200ul/well). Note: Each plate must have certain wells without cells as a blank control. o Blocking endogenous peroxidase: Incubate the fixed cells with 50ul (per well) of Pretreatment Solution (provided in the kit) for 10 minutes at room temperature and then rinse wells twice with PBS (200ul/well).

Procedure:

1. One-step Reaction: Mix 10ul (or 5-20ug) of primary antibody and 100ul of the 50X ELISA Probe (red solution provided in the kit) with 5ml Enhancer (blue solution provided in the kit) (this amount is sufficient for one 96-well plate). Add 50ul of this mixture to each well of the above fixed and pretreated cell plate. Incubate the plate for 60 minute at room temperature with a gentle shaking. Note: Each plate may needs certain wells incubated with Enhancer plus ELISA Probe only but without primary antibody as a negative control.

2. Wash: Wash all wells 3 times with 200ul of 1X Rapid Wash Solution (made by diluting 50ml of the kit-provided 10X Rapid Wash Solution with 450ml distilled water).

3. Development: To 10ml of distilled water, add 100ul of 100X T-buffer first, then, add 100ul of 100X TMB solution and mix to make 1XTMB solution. Apply 100ul of the 1X TMB solution to each well and wait till desired colors appear. Read the plate with a proper microplate reader and measure absorbance at 370nm (without stop) or at 450nm after stop the development by adding 100ul/well 1N HCL (or H2SO4).

Note: Reading at 450nm provides a better assay window than at 370nm. However, at 375nm without stopping user can continually reading plate till desired absorbance appear.

Related Products:

Cat# Product Name
WB-R51, WB-M51, WB-G51, WB-R50, WB-M50, WB-G50 UniversalTM One-step Western Blot Kits
IHC-R60, IHC-M60,IHC-G60, IHC-R61, IHC-M61, IHC-G61 UniversalTM One-step IHC Kits
E-R73, E-G73, E-R74, E-G74 UniversalTM One-step Sandwich ELISA Kits
E-R70, E-M70, E-G70, E-R71, E-M71, E-G71 UniversalTM One-step Indirect ELISA Kits
E-R90, E-M90, E-H90, E-G90 UniversalTM One-step IgG Quantification ELISA Kits