Attogene AU2044-01 Fluorescent Universal Human IgG IgM Lateral Flow Serology Kit User Manual

Attogene AU2044-01 Fluorescent Universal Human IgG IgM Lateral Flow

Serology Kit

Specifications

• Product Name: Fluorescent Universal Human IgG/IgM Lateral Flow Assay Kit
• Intended   Use: Laboratory Use
• Catalog   Number: AU2044-01
• For Research Use Only. Not for use in Diagnostic Procedures.

Product Usage Instructions

Introduction

• Attogene’s Fluorescent Human IgG/IgM universal lateral flow assay kit is a ready-to-use, universal test strip based on lateral flow technology.
• It uses quantum dot particles containing neutravidin to capture biotinylated antigens.
• The kit allows for qualitative or quantitative rapid test systems for the detection of anti-human IgG and IgM antibodies reacting to biotinylated antigens.

Kit Contents

• lateral flow dipsticks
• Assay Running Buffer
• Biotin Rabbit IgG (control line antigen 1ng/ul)
• Biotin Human IgG (IgG Control)
• Biotin Human IgM (IgM Control)
• Manual
• 96 Well Plate

Precautions

It is necessary to ensure the appropriate concentration ratio/stoichiometry between the biotinylated antigen and biotinylated rabbit IgG to observe test and control lines at the desired intensity.

Procedure

1. Prepare serially diluted biotinylated antigen in phosphate-buffered saline (e.g., 1ug/ml to 1ng/mL).
2. Prepare the clinical sample (e.g., human serum or plasma) or contrived sample (e.g., humanized IgG).
3. If higher analytical sensitivity is required, adjust the volume and/or concentration of the sample added into the well.

Interpretation of Results

The test contains lines dependent on antibody concentration in the sample. Adjusting the sample may be necessary to understand the dynamic range of the test.

What to expect:

• Test lines: Higher concentration = higher intensity.
• Control line: Intensity remains constant regardless of test line intensity.
• The drying time of strips affects line intensities; ensure proper drying time for accurate results.

Additional Analysis

Positive samples can be confirmed using nucleic acid analysis techniques like agarose or acrylamide gels. A fluorescent lateral flow reader can provide numerical readings.

FAQ

Q: How should I store the kit components?

• A: Refer to the storage instructions provided for each component in the kit contents list.
• Keep components refrigerated as needed.

Q: Can this kit be used for diagnostic purposes?

• A: No, this kit is for research use only and is not intended for diagnostic procedures.

Introduction

• Attogene’s Fluorescent Human IgG/IgM universal lateral flow assay kit is a ready-to-use, universal test strip, which is based on lateral flow technology that uses quantum dot particles containing neutravidin to conveniently capture biotinylated antigens.
• The device is designed to easily develop qualitative or quantitative rapid test systems for the detection of anti-human IgG and IgM antibodies that react to any antigen that can be biotinylated (i.e. viral antigen, autoimmune antigen) and is easily customizable providing every laboratory with the possibility to perform assay feasibility.
• Antibody tests are a method of choice to determine if a person has been exposed to a pathogen or not.
• They are also incredibly valuable in the detection of autoantibodies that can be found in human autoimmune disorders.
• In this test, a biotinylated antigen (User supplied) is mixed with a biotinylated rabbit IgG (binds to goat anti-rabbit control line) and the sample (human sera or plasma) is simply mixed into the specially designed assay running buffer in a well of the supplied 96-well plate, mixed and is then added to the sample port of the cassette. Generally, the reaction is complete in 10-15 minutes.
• It is very important to note that the relative stoichiometry between the biotinylated antigen, biotinylated rabbit IgG added, and the streptavidin gold is critical for assay optimization.
• The appropriate concentration of biotinylated antigen to use with strips is dependent upon the purity and sequence and a standard curve can be used to determine the relative ratio (generally between 1ng-100ng per test).
• A positive control line (biotin-rabbit IgG) antibody will bind to the goat anti-rabbit (GAR) line on the test to ensure the assay is running appropriately.

Features & Benefits

• Can be used for the development of a lateral flow assay for the detection of anti-human antibodies targeting the antigen of choice.
• No need to stripe-capture antibodies.
• The cost-effective way to screen for further downstream lateral flow assay development.

Kit Contents

Storage and Stability

• The kit should be stored at 2℃ – 30℃ until ready to use.
• The test must remain in the sealed container until use.

Required Materials Not Supplied

• Biotinylated antigen(s)
• Samples (contrived sample, human sera, or plasma)
• Centrifuge – For the preparation of clear specimens
• Pipettor and pipette tips – to transfer samples and controls.

Precautions

• Before use, ensure that the product has not expired by verifying that the date of use is before the expiration date on the label.
• The test strips are packaged in a vial that contains desiccant. Avoid prolonged periods of having the vial open to avoid desiccant becoming saturated.
• Avoid cross-contamination of samples by using a new tube and disposable pipette tip for each sample.
• Use only Lateral Flow Kit reagents from one kit lot, as they have been adjusted in combination.
• It is good laboratory practice to use positive and negative controls to ensure proper test performance.
• Due to the hook effects, if no signal is detected in the test line, a serial dilution may be necessary to bring the biotinylated antigen into the appropriate concentration ratio/stoichiometry with biotinylated rabbit IgG to see a test line and a control line at the desired intensity.

Procedure

Prepare serially diluted biotinylated antigen in phosphate-buffered saline (i.e., 1ug/ml to 1ng/mL). Prepare the clinical (e.g., human serum or plasma) or contrived (e.g., humanized IgG) sample.

Perform the following:

1. Add 100μL of Lateral Flow Running Buffer into a well of a 96-well plate.
2. Always run a positive and negative control well with a sample:
   • A. (TEST LINE REAGENTS) mix a designated amount (a volume 1μL-5μL are good starting point) of biotinylated antigen into the assay running buffer. When running an LFA for the first time, we recommend trying large dilutions of sample/antigen to determine the dynamic range of the assay and keeping the volume below 15μL if possible. As controls for the human IgG or human IgM capture lines, use 5ul of control biotin labeled human IgG or human IgM.
   • B. (CONTROL LINE REAGENT) mix 5μL of biotinylated rabbit IgG (5ng total)
   • C. (SAMPLE) Add between 1-10ul clinical (e.g., human serum or plasma) or contrived (e.g., humanized IgG) samples.
3. Mix each well completely by pipetting up and down several times.
4. Place test strip into the well and run vertically.
5. Incubate for 10-15 minutes.
6. Visually analyze the strip by eye in conjunction with an ultraviolet flashlight or photograph in a fluorescent strip reader.

• NOTE: If a higher analytical sensitivity is required, it could be helpful to increase the volume and/or concentration of the sample added into the well.
• Volume and concentration of analyte-specific solutions, and incubation time are always part of the individual test development.

Interpretation of Results

• This test is a lateral flow assay containing test lines that are dependent on the concentration of antibodies in the sample. Adjustment of the sample may be required to understand the dynamic range of the test.

What to expect at the test lines:

• The higher the concentration of human anti-IgG or IgM reactive against the biotinylated antigen in the sample the higher the intensity of the test line compared to the strip run in a mixture lacking sample (negative control strip).

What to expect at the control line:

• The intensity of the control will not decrease as the test lines increase.
• Determination made using strips that have dried for more or less than the required time may be inaccurate, as line intensities may vary with drying time.

Additional Analysis

• If necessary, positive samples can be confirmed by analyzing using a nucleic acid analysis technique such as agarose or acrylamide gels.
• A fluorescent lateral flow reader may also be employed to generate numerical readings.
• Contact Us 3913 Todd Lane, Suite 310
• Austin, TX 78744
• Phone: 512- 333-1330
• Email: sales@attogene.com
• Web: www.attogene.com
• AU2044-01_V1_20240112

Documents / Resources

| Attogene AU2044-01 Fluorescent Universal Human IgG IgM Lateral Flow Serology Kit [pdf] User Manual
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References

• Home Update - Attogene
• Home Update - Attogene

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