ELISA Starter Kit (Casein)

ELISA Starter Kit (Casein)

Catalog Number	K0331001
Size	1000well
Type	ELISA Kit
Datasheet

Specificity

This Kit is used to quantitatively measure levels of proteins or specific antigens in serum or other biological samples by using with EzWay ELISA Core kits. Except antibody & standard, all other reagents for full ELISA test are supplied.

Description

There is no need to buy extra solutions separately.
Minimize the cost and time for preparation of full ELISA Kit.
All kit components are optimized for common ELISA test.
It is convenient to use together with Core kits.

Kit Contents

Items	Quantity
ELISA Microtiter Wells Plate	10 plates
Coating Buffer (pH9.6)	125ml x 2
Blocking Solution (1% BSA/PBS)	125ml x 2
PBS Powder	Pouch for 1L x 5
Tween-20 (50%)	1ml (50%) x 5
TMB Solution	60ml
Substrate Solution (H2O2)	125ml
Stop Solution (2M H2SO4)	60ml

Storage & Shelf life

1 year at 2-8°C

Buffer Preparation

Coating Buffer: 50 mM Carbonate-Bicarbonate Buffer, pH 9.6. Resolve the coating material (antigen or antibody) in the coating buffer to make 1ug/ml (1-10ug/ml).
Sample/Standard/Antibody Diluting Solution : Dilute Sample/Standard/Antibody in PBS (Phosphate Buffered Saline, pH7.4). Or use PBST (Washing solution) or Blocking solution instead of PBS to help prevent non-specific binding.
Washing Solution: Add 50% 1ml Tween 20 to 1 Liter PBS and mix well.
Color Reaction Mixture: Mix 1 volume of TMB Solution and 2 volume of Substrate Solution (H2O2) prior to use.

Sandwich ELISA Protocol

Coating
1. Dispense 200ul (may vary from 50 to 200ul depending on user's need) of prepared Coating Solution to each well.
2. Incubate 1 hour at 37°C. (or 1-3 hours at room temperature/overnight at 4°C)
Washing (All washing method is the same)
1. Remove the solution from each well and fill up the Washing Solution. Repeat 3-5 times. Complete removal of liquid in each step is essential to good performance.
2. After the last wash, remove any remaining Washing Solution. Turn over the plate and blot carefully on paper towels.
Blocking
1. Add 250ul Blocking Solution to each well.
2. Incubate 1 hour at room temperature or at 37°C.
Washing
React Sample/Standard (or Primary Antibody)
1. Add 200ul diluted Sample/Standard (or Primary Antibody) to each well.
2. Incubate 1 hour at room temperature or at 37°C.
Washing
Add HRP-conjugated Detection Antibody (or Secondary Antibody)
1. Add 200ul diluted Detection Antibody to each well.
2. Incubate 1 hour at room temperature or at 37°C.
Washing
Color Reaction and Reading
1. Dispense 150ul Color Reaction Mixture into each well. (Or apply 50ul TMB Solution and 100ul Substrate Solution to each well. Mix well by vortexing or shaking slightly.)
2. After sufficient color development (5-10 minutes at room temperature or at 37°C), add 50ul Stop Solution (2M H2SO4) to each well.
3. Read plates in a microwell plate reader at wavelength setting of 450nm.