UniversalTM One-step IgG or IgE Quantification ELISA Kits (For R&D Use only)

UniversalTM One-step IgG or IgE Quantification ELISA Kits
I. Product Name, Catalog Number & Content:

Catalog #
Product Name
Size
 
Content
E-R90
UniversalTM One-step Rabbit IgG
Quantification ELISA Kit
480T (for 5 96-well plates)
 
10X Coating Buffer (10ML)
100XIgG Capture Ab-R (0.5ML)
Standard Rabbit IgG (5ug/ml, 100ul)
Enhancer (50ML)
100X ELISA Probe-R0 (0.5ML)
10X Rapid Wash Solution (50ML)
TMB Solution Set (for 100ML)
E-M90
UniversalTM One-step Mouse IgG
Quantification ELISA Kit
480T (for 5 96-well plates)
 
10X Coating Buffer (10ML)
100XIgG Capture Ab-M (0.5ML)
Standard Mouse IgG (5ug/ml, 100ul)
Enhancer (50ML)
50X ELISA Probe-M0 (0.5ML)
10X Rapid Wash Solution (50ML)
TMB Solution Set (for 100ML)
 
E-G90
UniversalTM One-step Goat IgG
Quantification ELISA Kit
480T (for 5 96-well plates)
 
10X Coating Buffer (10ML)
100XIgG Capture Ab-G (0.5ML)
Standard Goat IgG (5ug/ml, 100ul) Enhancer (50ML)
100X ELISA Probe-G0 (0.5ML)
10X Rapid Wash Solution (50ML)
TMB Solution Set (for 100ML)
E-H90
UniversalTM One-step Human IgG
Quantification ELISA Kit
480T (for 5 96-well plates)
 
10X Coating Buffer (10ML)
100XIgG Capture Ab-H (0.5ML) Standard Human IgG (5ug/ml, 100ul) Enhancer (50ML)
100X ELISA Probe-H0 (0.5ML)
10X Rapid Wash Solution (50ML)
TMB Solution Set (for 100ML)
E-hE90
UniversalTM One-step Human IgE
Quantification ELISA Kit
480T (for 5 96-well plates)
 
10X Coating Buffer (10ML)
100XIgE Capture Ab-hE (0.5ML) Standard Human IgE (5ug/ml, 100ul) Enhancer (50ML)
100X ELISA Probe-hE0 (0.5ML)
10X Rapid Wash Solution (50ML)
TMB Solution Set (for 100ML)
II. Product Description:

UniversalTM One-step IgG/E Detection ELISA is a latest breakthrough technology (patent pending) for identifying and quantifying total IgGs. Unlike regular

(classical) ELISA, the One-step IgG or IgE Detection ELISA combines blocking, IgG binding, washing and detector antibody binding into a rapid one-step

reaction. Therefore, using UniversalTM One-step IgG/E Detection ELISA kit, you can rapidly get your ELISA results around a couple of hours.



III. Application:

Detection and quantification of total IgG (including IgG antibodies) or IgE. Detection range: 0.5ng~50ng/ml or higher .

IV. Features:

Easy: One-step reaction instead of blocking, sample binding, washing and detector antibody binding.

Rapid: Get results around one hour (excluding coating time).

Reproducible: Highly reproducible results.

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

Capture antibody, Standard and ELISA Probe: store in -20 ≧ for up to 6 months or store in -70 ≧ manual defrost freezer for up to a year. Repeated freezing

and thawing is not recommended. Coating Buffer and Rapid Wash Solution: store in room temperature for up to one year. Note: Kit will be shipped in ambient temperature.

VI. Comparison:

One-step Procedure vs. Regular Procedure:

VII. Protocol:

i. Required materials but not included in the kit:

Stop solution (option): 1N HCl or 1N H2SO4. Assay Plate: 96-well high protein binding ELISA plate.

ii. Reagent Preparation:

1. Coating plate: For one 96-well plate, dilute 10ul of the 100X capture antibody with 10ml of 1x coating buffer (both are provided in the kit). Apply

100ul of this coating solution to each well and incubate for 1-2 hours at room temperature (or overnight at 4oC). Remove the coating solution and wash wells

three times with 200ul/well of 1X Rapid Wash Solution (provided in the kit).

2. 1X ELISA Probe solution: Dilute 100ul of the 100X ELISA Probe (the red solution provided in the kit) with 10ml Enhancer (the blue solution provided

in the kit) and mix well. This amount is sufficient for one 96-well plate.

3. 10X Serial diluted Standard solution:

For detection range 1~100 ng/ml: Prepare 8 tubes and mark the tubes with #1~ #8. To tube#1, add 32ul and to tube #2~#8, add 20ul of PBS buffer (or the

same buffer used for samples to be tested). Transfer 8ul of the 5ug/ml standard (provided in the kit) to tube #1 and mix well. Transfer 20ul of the standard

solution from tube #1 to tube #2 and mix. Transfer 20ul of the diluted standard solution from tube #2 to tube #3 and mix. Repeat such dilution to make the

following 10X serial dilutions: 1000 ng/ml (tube #1), 500 ng/ml (tube #2), 250ng/ml (tube #3), 125ng/ml (tube #4), 62.5 ng/ml (tube #5), 31ng/ml (tube #6),

16 ng/ml (tube #7) and 8ng/ml (tube #8) respectively.

For detection range 0.1~10 ng/ml: Prepare 8 tubes and mark the tubes with #1~ #8. To tube#1, add 196ul and to tube #2~#8, add 100ul of PBS buffer (or

the same buffer used for samples to be tested). Transfer 8ul of the 5ug/ml standard (provided in the kit) to tube #1 and mix well. Transfer 100ul of the

standard solution from tube #1 to tube #2 and mix. Transfer 100ul of the diluted standard solution from tube #2 to tube #3 and mix. Repeat such dilution to

make the following 10X serial dilutions: 100 ng/ml (tube #1), 50 ng/ml (tube #2), 25ng/ml (tube #3), 12.5ng/ml (tube #4), 6.25 ng/ml (tube #5), 3.1ng/ml

(tube #6), 1.6 ng/ml (tube #7) and 0.8ng/ml (tube #8) respectively.

iii. Procedure:

1. One-step Reaction: Add 90ul of the above pre-diluted 1XELISA probe/Enhancer solution to each well of the capture- antibody-coated ELISA plate, then

add 10ul/well of the serial diluted 10Xstandard to wells A1~H1(or A1,A2~H1,H2 for duplicate tests), add 10ul/well of samples to be tested to other wells (for

duplicate tests make two wells for each sample). Note: make at least two wells as negative controls for each plate by add 10ul buffer. Incubate the plate

for 50 minute at room temperature with gently shaking.

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). Empty the wells.

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 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

standard curve appear.

iv. Example:

Standard curve generated by Universal One-step IgG Quantification ELISA Kit (E-M90):

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-R80, E-M80, E-G80, E-R81,
E-M81, E-G81
UniversalTM One-step Cell-based ELISA Kits
E-R70, E-M70, E-G70, E-R71,
E-M71, E-G71
UniversalTM One-step Indirect ELISA Kits