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FlexLISA® Kits

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FlexLISA® Kits

Flexible, quick and easy ELISA development and optimization!

The Innova Biosciences FlexLISA® kits are designed for the development and optimization of sandwich ELISA assays to detect the presence of any antigen with a high affinity for any antibody pair in complex samples (serum, plasma, urine etc.).

Benefits of the FlexLISA® kit:

• Easy antibody labeling – uses world leading Lightning-Link® technology

• Rapid

• Uses 40 times less capture antibody – save on cost of materials

• Use your antibodies of choice

• Conjugate up to 3 capture and 3 detection antibodies – ideal for screening

• Flexibility to run multiple conjugates on a plate simultaneously

• Choice of HRP or AP for enzymatic detection

 

FlexLISA ELISA assay kit

A rapid one-step protocol is used in this kit, employing pre-blocked microtitre well strips coated with biotin. The FlexLISA® assay is run by adding the sample and the antibody mix to the wells, incubating for 1 hour, washing and reading the assay plate. FlexLISA® is ideal for quick and reliable ELISA assay development, antibody pair screening and ELISA assay optimization.

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Data

FlexLISA Figure 1. A FlexLISA Figure 1. B

FlexLISA Figure 1. c

Figure 1. Example of ELISA assay for the detection of CRP using LL-HRP to conjugate the detection antibody and making a calibration curve using CRP standards. Detection reagent: TMB.

Standard curves fitted with a linear equation (A) and a 4-parameter logistic equation (B). Intra-assay % CV (n=24) have been determined for 800, 200 and 50pg/ml CRP (C).

 

FlexLISA Figure 2 for website

Figure 2. Example of ELISA assay for the detection of CRP using LL-HRP and Ampliflu® Red as detection reagent (Excitation wavelength: 530nm, Emission wavelength: 590nm)

FlexLISA Figure 3. A FlexLISA Figure 3. B

FlexLISA Figure 3. c

 

Figure 3. Example of ELISA assay for the detection of CRP using LL-AP to conjugate the detection antibody and making a calibration curve using CRP standards. Detection reagent: PNPP. Standard curves fitted with a linear equation (A) and a 4-parameter logistic equation (B). Intra-assay % CV (n=24) have been determined for 800, 200 and 50pg/ml CRP (C).

 FlexLISA Figure 4 fo website

Figure 4. CRP depleted human serum was spiked with 250pg/ml CRP and serial dilutions of serum were made and loaded to a clear 96 stripwell plate in presence of anti CRP-streptavidin capture antibody and anti CRP-HRP or AP detection antibody. A standard curve using known amounts of CRP was also loaded to the plate. HRP and AP were detected using TMB and PNPP respectively. Once the standard curve had been fitted, the CRP concentrations of the samples added to the plate were determined by interpolating the measured absorbance. For all dilutions, the calculated CRP amount was within 10% the real value.

Kit components

The kit contains:

  • Lightning-Link® conjugation technology for the labeling of the capture and detection antibodies of choice
  • A biotin pre-coated 96 well stripwell plate which can be provided either as clear or black, depending on the enzymatic substrate (12 x 8 -strip well) in a re-sealable foil pouch

Not provided: antibodies, assay buffers, enzyme detection solutions

*Each 10ug vial is enough to run a 96 well plate: less than 0.1ug of capture or detection antibody/well is required and the 3 reactions provide the flexibility to run multiple conjugates/assays for optimization

FAQs

1. What is different about the FlexLISA® kit?

In a standard ELISA the capture antibody is bound directly to the surface of the well, whereas the 96 stripwell plate provided within the FlexLISA® kit is pre-coated with biotin which binds the capture antibody-streptavidin conjugate with a strong affinity. Therefore, much less capture antibody (40x less) is required compared to standard ELISA assays, saving on costs of expensive antibody.

Unlike other ELISA kits available on the market, the FlexLISA® kit also gives the flexibility of being able to use any antibody of your choice and conjugate as many capture and detection antibodies as required, giving complete flexibility.

Furthermore, the FlexLISA kit contains Innova Biosciences’ Lightning-Link®, the world leading antibody labeling technology, for covalently conjugating your capture and detection antibody. Lightning-Link® kits enable conjugation of any antibody with a free lysine, and each Lightning-Link® kit contains 3 reactions, providing the flexibility to run multiple conjugates at the same time.

 

2. Are all materials required to run the FlexLISA® included in the kit?

The FlexLISA® kit contains a 96 well plate and conjugation kits to label 3 different capture antibodies and 3 different detection antibodies. Antibodies, assay buffers and enzyme detection solutions are not provided, please see the protocol for additional materials required.

 

3. How much label and antibody is required for this kit?

The 3x10ug Lightning-Link® labeling kits provided within the FlexLISA® kit will each conjugate 10ug of antibody which is enough to run the 96 well plate. Less than 0.1ug of capture and detection antibody is required per well and the 3 reactions provide the flexibility to run multiple conjugates at once.

 

4. Do I need to block the plate?

The 96 well stripwell plates are pre-coated with biotin and pre-blocked with blocking agents to prevent high background.

 

5. What if my sample contains biotin?

The assay can tolerate presence of biotin up to 200pg/ml in the well. If biotin levels in your sample are higher, perform a 1 hour pre-incubation with the Ab mix and then add the sample directly to the plate (no need for washing step).

 

6. Which substrates do you recommend?

The substrate required depends on the choice of detection label. For HRP the most common substrates are TMB, QuantaRed™ QuantaBlu™, AmpliFlu™ Red, for Alkaline Phosphatase, PNPP, 4-MUP are common place.

 

7. How should I store my FlexLISA kit?

The kits are shipped at ambient temperature, upon receipt make sure to store at -20°C. All the buffers provided within the kit can be stored at either 4°C or -20°C.

 

8. How can I avoid high background in my assay?

Please see the below table for possible causes of high background and recommended solutions.

Problem

Possible Cause

Recommended Solution

 

 

High background

  

 

Insufficient washing

Increase the number of wash steps

Insufficient sample dilution

Increase sample dilution

Insufficient blocking

Increase blocking agent concentration in the assay buffer

Contaminated detection solution

Repeat assay

 

9. I am getting low signal in my assay, what shall I do?

Please see the below table for possible causes of signal and recommended solutions.

Problem

Possible Cause

Recommended Solution

 

Low Signal

 

 

 

Insufficient assay incubation time

Increase assay incubation time to 2 - 5 hours

Insufficient substrate solution incubation time

Check substrate solution detection protocol (supplier)

Incorrect plate reader settings

Check settings used

 

10. What should I do about variable replicates?

Please see the below table for possible causes of variable replicates and recommended solutions.

Problem

Possible Cause

Recommended Solution

 

Variable Replicates

  

Poor pipetting technique

Check against best practice. We advise reverse pipetting for standard, sample and Antibody Mix loading

Contamination of samples during addition to plate

Check against best practice

Blocked plate washer

Check all plate washer channels

 

Buy Online

FlexLISA HRP 1 x 96 well clear stripwell plate (4200-0010)

In stock

£350.00

FlexLISA AP 1 x 96 well clear stripwell plate (4200-0020)

In stock

£350.00

FlexLISA HRP 1 x 96 well black stripwell plate (4200-0030)

In stock

£350.00

FlexLISA AP 1 x 96 well black stripwell plate (4200-0040)

In stock

£350.00
Our shipping times are as follows:
48 hours to the US. 24 to 48 hours to Europe.
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