Amplite® Human Apolipoprotein A1 (ApoA1) Kit *Optimized For ELISA Development with HRP*
Ordering information
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Storage, safety and handling
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12171501 |
Components
Example protocol
AT A GLANCE
AT A GLANCE
Intended use: For quantitative determination of human Apolipoprotein A1 (apoA1) in serum/ plasma samples and cell culture supernatants. Please note that wash-, block- and incubation buffers should contain detergent. Tween 20, Triton X-100 or NP40 can be used at a concentration of 0.05-0.5%. In block and incubation buffers it is recommended to use 0.1% BSA, but not bovine serum, as HDL 44 also binds bovine apoA1.
Serum/plasma samples: When analyzing human serum/plasma samples it is recommended to use Apo ELISA buffer for dilution of samples, standard and detection an- tibody. The buffer prevents false positive read-outs which may be caused by interference of hetero- philic antibodies commonly found in human plasma and serum. Triton X-treatment of samples, necessary for apoB analysis, will not interfere with apoA1 analysis. It is recommended to dilute se- rum/plasma samples 150,000x to 200,000x. Avoid repeated freezing-thawing cycles and do not store samples in -20°C. Samples stored in -20°C will give false high apoA1 values.
Reagents: Antibodies are supplied in sterile-filtered (0.2 μm) PBS with sodium azide (0.02%). Streptavidin-HRP is supplied in PBS with 1% BSA and 0.002% Kathon CG.
Standard range: 0.6-40 ng/ml
Intended use: For quantitative determination of human Apolipoprotein A1 (apoA1) in serum/ plasma samples and cell culture supernatants. Please note that wash-, block- and incubation buffers should contain detergent. Tween 20, Triton X-100 or NP40 can be used at a concentration of 0.05-0.5%. In block and incubation buffers it is recommended to use 0.1% BSA, but not bovine serum, as HDL 44 also binds bovine apoA1.
Serum/plasma samples: When analyzing human serum/plasma samples it is recommended to use Apo ELISA buffer for dilution of samples, standard and detection an- tibody. The buffer prevents false positive read-outs which may be caused by interference of hetero- philic antibodies commonly found in human plasma and serum. Triton X-treatment of samples, necessary for apoB analysis, will not interfere with apoA1 analysis. It is recommended to dilute se- rum/plasma samples 150,000x to 200,000x. Avoid repeated freezing-thawing cycles and do not store samples in -20°C. Samples stored in -20°C will give false high apoA1 values.
Reagents: Antibodies are supplied in sterile-filtered (0.2 μm) PBS with sodium azide (0.02%). Streptavidin-HRP is supplied in PBS with 1% BSA and 0.002% Kathon CG.
Standard range: 0.6-40 ng/ml
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
Prepare apoA1 standard by reconstituting contents of vial 4 in 1 ml PBS with 1% BSA, do not stir and leave at room temperature for 15 minutes followed by vortex for 3 sek. This gives a stock solution of 4 μg/ml which should be used immediately or stored in aliquots at -20°C for future use. We recommend the aliquots not to be refrozen after initial use. For the test, prepare dilutions of the stock using the standard range as a guideline.
Prepare apoA1 standard by reconstituting contents of vial 4 in 1 ml PBS with 1% BSA, do not stir and leave at room temperature for 15 minutes followed by vortex for 3 sek. This gives a stock solution of 4 μg/ml which should be used immediately or stored in aliquots at -20°C for future use. We recommend the aliquots not to be refrozen after initial use. For the test, prepare dilutions of the stock using the standard range as a guideline.
SAMPLE EXPERIMENTAL PROTOCOL
- Coat a high protein binding ELISA plate with mAb HDL 110, diluted to 2 μg/ml in PBS, pH 7.4, by adding 100 μl/well. Incubate overnight at 4-8°C.
- Wash twice with PBS (200 μl/well).
- Block plate by adding 200 μl/well of PBS with 0.05% Tween 20 containing 0.1% BSA (incubation buffer). Incubate for 1 hour at room temperature.
- Wash five times with PBS containing 0.05% Tween.
- Prepare apoA1 standard by reconstituting contents of vial 4 in 1 ml PBS with 1% BSA, do not stir and leave at room temperature for 15 minutes followed by vortex for 3 sek. This gives a stock solution of 4 μg/ml which should be used immediately or stored in aliquots at -20°C for future use. We recommend the aliquots not to be refrozen after initial use. For the test, prepare dilutions of the stock using the stand- ard range as a guideline.
- Prepare apoA1 standard by reconstituting contents of vial 4 in 1 ml PBS with 1% BSA, do not stir and leave at room temperature for 15 minutes followed by vortex for 3 sek. This gives a stock solution of 4 μg/ml which should be used immediately or stored in aliquots at -20°C for future use. We recommend the aliquots not to be refrozen after initial use. For the test, prepare dilutions of the stock using the stand- ard range as a guideline.
- Wash as in step 4.
- Add 100 μl/well of mAb HDL 44-biotin at 0.5 μg/ml in incubation buffer or Assay buffer for serum/plasma samples. Incubate for 1 hour at room temperature.
- Wash as in step 4.
- Add 100 μl/well of Streptavidin-HRP diluted 1:1000 in incubation buffer. Incubate for 1 hour at room temperature. Please note that sodium azide used in buffers will inhibit HRP activity.
- Wash as in step 4.
- Add 100 μl/well of appropriate substrate solution e.g. TMB, available from Mabtech product code 3652-F10.
- Measure the optical density in an ELISA reader after suitable developing time. If required stop the reaction first.
Application notes
A Meta-Analysis of Common Calcium Indicators
A New Protein Crosslinking Method for Labeling and Modifying Antibodies
A Novel Fluorescent Probe for Imaging and Detecting Hydroxyl Radical in Living Cells
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
Abbreviation of Common Chemical Compounds Related to Peptides
A New Protein Crosslinking Method for Labeling and Modifying Antibodies
A Novel Fluorescent Probe for Imaging and Detecting Hydroxyl Radical in Living Cells
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
Abbreviation of Common Chemical Compounds Related to Peptides
FAQ
Are Cell Navigator® Cell Plasma Membrane Staining Kits suitable for cell culture medium samples?
Are there any alternatives to BrdU (Bromodeoxyuridine)?
Are there any alternatives to Cy5?
Are there any alternatives to indocyanine green (ICG)?
Are there any calcium indicators that don't require probenecid (PBC)?
Are there any alternatives to BrdU (Bromodeoxyuridine)?
Are there any alternatives to Cy5?
Are there any alternatives to indocyanine green (ICG)?
Are there any calcium indicators that don't require probenecid (PBC)?