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Test Results

  • Test Results
  • Compatibility
  • Batch to Batch

Compatible with many reagents, low detection limit, high dynamic range

Tested R&D Systems DuoSet® ELISA

No. Tested DuoSet name Catalog code
1 Human CXCL10/IP-10 DY266
2 Human GDNF DY212
3 Human IFN-gamma DY285
4 Human IL-2 DY202
5 Human IL-6 DY206
6 Human IL-8/CXCL8 DY210
7 Human TNF-alpha DY208
8 Human VEGF DY293B
9 HIV-1 Gag p24 DY7360-05
10 Rat TIM-1/KIM-1/HAVCR DY3689
11 Rat VEGF DY564
12 Human IFN-alpha DY9345-05
13 Human IL-10 DY217B
14 Human IL-15 DY247
15 Human TGF-beta 1 DY240
16 Human C-Reactive Protein/CRP DY1707
  • Same or better detection limit
  • ~10X larger dynamic range

Substrate: QuantaRed™ Enhanced Chemifluorescent HRP Substrate Kit from Thermo Fisher

Example 1 (IL-6 and TNF-a)

MicroFluere®
IL-6

  • Time saving: <1 hour vs. 6 hours
  • Reagent cost saving: 75%

MicroFluere®
TNF-a

Example 2 (Rat VEGF)

MicroFluere® plate
Fluorescence measurement

  • R-square: 0.99867
  • LOD: 12.5 pg/mL
  • Dynamic range: 12.5-9,000 pg/mL
  • Sample/reagent volume: ≤20 µL
  • Assay time: 40 minutes

Traditional plate
Colorimetric measurement

  • Dynamic range: 15.6-1,000 pg/mL
  • Sample/reagent volume: ≥100 µL
  • Assay time: 280 minutes

*https://www.rndsystems.com/products/rat-vegf-duoset-elisa_dy564

Example 3 (Rat TIM-1/KIM-1/HAVCR)

MicroFluere® plate
Fluorescence measurement

  • R-square: 0.99081
  • LOD: 11.3 pg/mL
  • Dynamic range: 11.3-9,000 pg/mL
  • Sample/reagent volume: ≤20 µL
  • Assay time: 40 minutes

Traditional plate
Fluorescence measurement

  • R-square: 0.99948
  • LOD: 10.2 pg/mL
  • Dynamic range: 10.2-8000 pg/mL
  • Sample/reagent volume: : ≥100 µL
  • Assay time: 270 minutes

Traditional plate
Colorimetric measurement

  • Dynamic range: 15.6-1,000 pg/mL
  • Sample/reagent volume: ≥100 µL
  • Assay time: 280 minutes

*https://www.rndsystems.com/products/rat-vegf-duoset-elisa_dy3689

Example 4 (p24 detection)

Results from the University of Missouri using traditional 96-well plate. Very poor linearity. No three points can form a line.

Results from the same group at the University of Missouri using R&D Systems ELISA kit and MicroFluere® 96-well plate. Excellent linearity.

Example of Competitive ELISA

  • Collaborative work with University of Michigan
Traditional well plate MicroFluere
Coating and Blocking  Over 1,000 minutes 100 minutes
Detection protocol Over 120 minutes 20 minutes
Sample/reagent volume 100 mL ≤20 mL

Example of Immunoglobulin G Detection

  • Collaborative work with Shanghai ZhenGe Biotech Co, LTD. (Shanghai, China)
Traditional well plate MicroFluere
Coating and Blocking  Over 1,000 minutes 90 minutes
Detection protocol Over 150 minutes 30 minutes
Sample/reagent volume 100 mL ≤20 mL

Example of Antibody Detection

  • Collaborative work with PUMC Hospital (Beijing, China)
Traditional well plate MicroFluere
Coating and Blocking  Over 2,000 minutes 90 minutes
Detection protocol Over 100 minutes 25 minutes
Sample/reagent volume 100 mL ≤20 mL

COVID-19 related

  • N protein in serum
  • LOD: 20 pg/mL
  • Assay time: 30 minutes
  • IgG in serum
  • LOD: ~1 ng/mL (not optimized yet)
  • Assay time: 25 minutes
  • Throughput: 80 samples + 16 standards. In-experiment standards, more accurate
  • All detection is done using 20 µL of sample and reagents

Compatibility tested commercial ELISA readers

  1. EnSpire 2300 Multimode Plate Reader (PerkinElmer)
  2. Infinite F200 Fluorescent Microplate Reader System (Tecan)
  3. Synergy Neo2 Hybrid Multi-Mode Reader (BioTek Instruments)
  4. Synergy H1 Hybrid Multi-Mode Reader (BioTek Instruments)
  5. Synergy HT (BioTek Instruments)
  6. GloMax®-Multi+ Detection System (Promega)
  7. Varioskan Flash (Thermo Fisher Scientific)

Intra- assay CVs

 

Plate 1

Conc. (pg/mL)

Rep 1

Rep 2

Rep 3

Avg

STD

CV%

0

399

365

412

392

24

6.2%

3.125

511

464

464

480

27

5.7%

6.25

585

550

461

532

64

12.0%

12.5

661

667

704

677

23

3.4%

25

923

1027

911

954

64

6.7%

50

1452

1510

1536

1499

43

2.9%

100

2287

2619

2607

2504

188

7.5%

200

4638

4672

4234

4515

244

5.4%

Plate 2

Conc. (pg/mL)

Rep A

Rep B

Rep C

Avg

STD

CV%

0

450

379

369

399

44

11.1%

3.125

454

439

401

431

27

6.3%

6.25

592

471

474

512

69

13.5%

12.5

763

587

680

677

88

13.0%

25

1008

993

872

958

75

7.8%

50

1710

1581

1514

1602

100

6.2%

100

2957

2334

2665

2652

312

11.8%

200

4702

4807

5020

4843

162

3.3%

Inter- assay CVs

Conc. (pg/mL)

Plate 1

Plate 2

Avg

STD

CV%

Rep 1

Rep 2

Rep 3

Rep A

Rep B

Rep C

0

399

365

412

450

379

369

396

32

8.1%

3.125

511

464

464

454

439

401

456

36

7.9%

6.25

585

550

461

592

471

474

522

60

11.6%

12.5

661

667

704

763

587

680

677

58

8.5%

25

923

1027

911

1008

993

872

956

62

6.5%

50

1452

1510

1536

1710

1581

1514

1551

89

5.7%

100

2287

2619

2607

2957

2334

2665

2578

244

9.5%

200

4638

4672

4234

4702

4807

5020

4679

258

5.5%