Intended research use:

  • Studies indicate that liver deposits of Collagen IV, a type of collagen found primarily in basement membrane, have been found to be increased with liver fibrosis.  Furthermore, studies show that this increased liver Collagen IV directly correlates with an increase in serum Collagen IV.
  • Designed for research purposes only, not intended for diagnostic purposes.
  • Biomarker in research of fibrotic conditions often associated with liver-disease (NAFLD/NASH and drug induced liver injury).
  • For studies related to chronic liver injury.  Assay is complimentary to CK18, HA and α–GST ELISAs, making it well-suited for a biomarker panel researching drug-induced liver injury (DILI).
  • Complete ELISA kit
  • Standardized and quantitative
  • Only 50 µl of sample is needed
  • Ready-to-use pre-coated ELISA plates

The Diapharma Serum Collagen IV ELISA is designed for the assay of serum collagen IV. It is a solid phase one-step sandwich ELISA. Collagen IV in the sample is bound simultaneously by a solid-phase monoclonal antibody and a monoclonal antibody-enzyme conjugate, each directed at different antigenic sites. The monoclonal antibodies react to the 7S domain and the triple helix domain of human placental type IV collagen.  This results in the collagen IV molecule being sandwiched between the solid phase and enzyme-labelled antibodies. After removing unbound enzyme-labeled antibody and sample, the plate is incubated with enzyme substrate. The resultant color development is directly proportional to the amount of collagen IV in the sample.

Typical Standard Curve

serum collagen elisa assay measurement

Typical calibration curve obtained using Serum Collagen IV ELISA. 4-Parameter plot of A450/630 nm versus ug/L Collagen IV. Assay range is 15.6-1000 ug/L Collagen IV.

Details

Availability / Lead Time:  This kit is manufactured at the time of order and may have a 3 month production time depending on stock status.  Minimum order quantity is 20 kits/order.

Manufactured by:  Manufactured for Diapharma Group

Detection Range:  15.6-1000 µg/mL

Sensitivity:  15.6 µg/mL

Sample volume:  50 µL

Sample type:  Serum

Incubation time:  1 hour

Format:  Sandwich ELISA

Reference values:  99 ± 23 µg/L (Mean ± 1 SD, N=180)
It is recommended that each laboratory establish its own reference range.

Species:  Human; other species may be cross-reactive, but no formal testing has been completed

Cross reaction:  Cross reactivity is less than 2% with Collagen II and less than 0.5% with other forms of collagen.

Sensitivity:  When reading from the standard curve the A450nm value of the 1000 µg/L standard should be >0.6 OD.

  • Collagen IV Antibody coated Microassay Plate: 12 strips of 8 wells (96 break apart wells in total), in a frame. Ready to Use.
  • Collagen IV Calibrator:  Stock Solution, 1000 ug/L
  • 10X Wash Buffer Concentrate
  • Dilution Buffer:  Ready to Use
  • Enzyme Conjugate: Anti-collagen IV mouse Fab’ conjugated to HRP, Ready to Use
  • TMB Substrate: Ready to Use
  • Stop Solution: 1 M Sulfuric acid, Ready to Use
  • Uncoated Microassay: Plate Ready to Use

 

Interference: 

No significant interference has been observed in this assay with lipemic, hemolytic or icteric samples.

  • Lipemia: Less than 10% interference up to 1200 Formazin turbidity units.
  • Hemolysis: Less than 10% interference up to 3 g/L hemoglobin.
  • Icterus: Less than 10% interference up to 0.2 g/L bilirubin.

Dilution Recovery:

Dilution of samples containing high levels of collagen IV gave the following results:

Sample Dilution
1 / 2 1 / 4 1 / 8
Expected µg/L Obtained µg/L Recovery % Expected µg/L Obtained µg/L Recovery % Expected μg/L Obtained µg/L Recovery %
A 57 61 107 28 32 114 14 16 114
B 107 110 103 53 58 109 27 28 104
C 259 270 104 130 139 107 65 67 103

 

Reproducibility:

Intra-assay variation of the DIAPHARMA Serum Collagen IV ELISA.

Sample [Collagen IV]

µg/L

SD %CV N
Low 119 7.4 6.2 8
Medium 218 7.8 3.6 8
High 520 12 2.3 8

 

Inter-assay variation:

Intra-assay variation of the DIAPHARMA Serum Collagen IV ELISA.

 

Sample Collagen IV]

µg/L

SD %CV N
Low 115 11 9.6 6
Medium 291 13 4.5 6
High 370 31 8.2 6

 

Inter-batch variation:

Inter-batch variation of the DIAPHARMA Serum Collagen IV ELISA calculated for three batches of kits.

 

Sample Collagen IV]

µg/L

SD %CV N
Low 115 5 4.3 3
Medium 270 6 2.2 3
High 386 16 4.1 3

 

Collagen IV is a type of non-helical forming collagen found primarily in the basal lamina (basement membrane).

Chronic liver disease comprises a number of progressive disorders which culminate in liver cirrhosis and which are characterized by excessive deposition of collagen.

Although various types of collagen (type I, III, IV, V and VI) increase in the liver with the progression of fibrosis, type IV collagen, a constituent of the basement membrane, is particularly noteworthy for the following reasons: its serum level correlates with hepatic levels of collagen IV, serum levels of collagen IV fall in response to potential therapies being developed and it is the earliest type of collagen to be synthesized during experimental liver injury. Serum collagen IV levels are elevated in a variety of liver diseases; in particular, serum collagen levels may be used to study potential new response in Hepatitis C infection, and in abstaining alcoholics.