Human Apolipoprotein A1
ELISA development kit
Product Code: 3710-1H-20

CONTENTS, development kit for 20 plates:
Vial 1 (green top)
Monoclonal antibody HDL 110 (500 μl)
Concentration: 1 mg/ml
Vial 2 (yellow top)
Biotinylated monoclonal antibody HDL 44 (250 μl)
Concentration: 1 mg/ml
Vial 3 (white top)
Streptavidin-Horseradish Peroxidase (250 μl)
Vial 4
Lyophilised purified apoA1 standard batch 9 (4 μg)
To ensure total recovery of stated quantity, vials have been overfilled.
STORAGE:
Shipped at ambient temperature. On arrival box 1 should be stored
refrigerated at 4-8°C and box 2 should be stored frozen at -20°C.

GeneralIntended 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 isrecommended 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 (product code: 3652-M2) for dilution of samples, standard and detection antibody. The buffer prevents false positive read-outs which may be caused by interference of heterophilic 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 serum/plasma samples 150,000x to 200,000x, see dilution guidelines at https://www.mabtech.com/
knowledge-center/apodilution. 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
Standard calibration: The standard has been calibrated against an international standard fromWHO. One ng of supplied standard equals one ng of WHO-IFCC:SP1-01 standard. Please notethat the calibration is batch specific.

OD 450 nm

10.0

1.0

0.10.1

110Concentration of apoA1 standard (ng/ml)

Guidelines for Human Apolipoprotein A1 ELISADay 1 1.

Coat a high protein binding ELISA plate with mAb HDL 110, diluted to 2 μg/ml inPBS, pH 7.4, by adding 100 μl/well. Incubate overnight at 4-8°C.

Day 2 2.

Wash twice with PBS (200 μl/well).

3.

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.

4.

Wash five times with PBS containing 0.05% Tween.

5.

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 berefrozen after initial use. For the test, prepare dilutions of the stock using the standard range as a guideline.

6.

Add 100 μl/well of samples or standards diluted in incubation buffer or Apo ELISAbuffer for serum/plasma samples and incubate for 1 to 2 hours at room temperature.
Dilution recommendations for serum/plasma samples can be found at https://www.
mabtech.com/knowledge-center/apodilution.

7.

Wash as in step 4.

8.

Add 100 μl/well of mAb HDL 44-biotin at 0.5 μg/ml in incubation buffer or Apo
ELISA buffer for serum/plasma samples. Incubate for 1 hour at room temperature.

9.

Wash as in step 4.

10. Add 100 μl/well of Streptavidin-HRP diluted 1:1000 in incubation buffer. Incubatefor 1 hour at room temperature. Please note that sodium azide used in buffers will
inhibit HRP activity.
11. Wash as in step 4.
12. Add 100 μl/well of appropriate substrate solution e.g. TMB, available from Mabtech
product code 3652-F10.
13. Measure the optical density in an ELISA reader after suitable developing time. If
required stop the reaction first.

Updated on 2020/03/31

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Developed and manufactured by MABTECH AB, Sweden,
whose quality management system complies with thestandards ISO 9001:2015 & ISO 13485:2016.

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The products are for research use only.
MABTECH shall not be liable for the use or handling of the product or for consequential,special, indirect or incidental damages there from.
Mabtech AB (Head Office)SwedenTel: +46 8 716 27 00
mabtech@mabtech.com

www.mabtech.com

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USA
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mabtech.usa@mabtech.com