Product datasheet

Human Pro-Collagen I alpha 1 ELISA Kit ab210966

11 References 6 Images

Overview

Product name Human Pro-Collagen I alpha 1 ELISA Kit

Detection method Colorimetric
Precision Intra-assay

Sample n Mean SD CV%

Serum 8 1.8%

Inter-assay

Sample n Mean SD CV%

Serum 3 3%

Sample type Cell culture supernatant, Serum, Plasma, Cell culture extracts, Tissue Extracts
Assay type Sandwich (quantitative)
Sensitivity 5.3 pg/ml
Range 39.06 pg/ml - 2500 pg/ml
Recovery Sample specific recovery

Sample type Average % Range

Serum 93 91% - 94%

Cell culture media 99 97% - 101%

Hep Plasma 101 94% - 107%

EDTA Plasma 108 105% - 114%

Cit plasma 106 102% - 110%

Assay time 1h 30m

1
Assay duration One step assay
Species reactivity Reacts with: Human
Does not react with: Cow
Product overview Pro-Collagen I alpha 1 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is
designed for the quantitative measurement of human Pro-Collagen I alpha 1 protein in serum,
plasma, cell culture supernatants, and cell and tissue extract samples.

The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter
conjugated detector antibody which immunocapture the sample analyte in solution. This entire
complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of
an anti-tag antibody coating the well. To perform the assay, samples or standards are added to
the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound
material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue
coloration. This reaction is then stopped by addition of Stop Solution completing any color change
from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the
intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB
can be recorded kinetically at 600 nm.

Sensitivity:
Samples diluted in Sample Diluent NS MDD = 5.6 pg/mL
Samples diluted in 1X Cell Extraction Buffer PTR MDD = 5.3 pg/mL

Notes Type I collagen is the most abundant structural protein of connective tissues such as skin, bone
and tendon. It is synthesized as a pro-collagen molecule that is characterized by a 300 nm triple
helical domain flanked by globular N- and C-terminal propeptides. Specifically, human Pro-
Collagen I alpha 1 consists of a signal peptide (amino acids (aa) 1-22), a propeptide (aa 23-161),
the mature chain (aa 162-1218), and another propeptide (aa 1219 – 1464). The non-helical
propeptides are removed by procollagen N- and C-proteinase activities so that the mature triple
helices can self-assemble into collagen fibrils that provide tensile strength to tissues.

Platform Pre-coated microplate (12 x 8 well strips)

Properties

Storage instructions Store at +4°C. Please refer to protocols.

Components 1 x 96 tests

10X Human Pro-Collagen I alpha 1 Capture Antibody 1 x 600µl

10X Human Pro-Collagen I alpha 1 Detector Antibody 1 x 600µl

10X Wash Buffer PT (ab206977) 1 x 20ml

50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml

5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml

Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml

Human Pro-Collagen I alpha 1 Lyophilized Recombinant Protein 2 vials

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Components 1 x 96 tests

Plate Seals 1 unit

Sample Diluent NS (ab193972) 1 x 50ml

SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit

Stop Solution 1 x 12ml

TMB Development Solution 1 x 12ml

Function Type I collagen is a member of group I collagen (fibrillar forming collagen).

Tissue specificity Forms the fibrils of tendon, ligaments and bones. In bones the fibrils are mineralized with calcium
hydroxyapatite.
Involvement in disease Defects in COL1A1 are the cause of Caffey disease (CAFFD) [MIM:114000]; also known as
infantile cortical hyperostosis. Caffey disease is characterized by an infantile episode of massive
subperiosteal new bone formation that typically involves the diaphyses of the long bones,
mandible, and clavicles. The involved bones may also appear inflamed, with painful swelling and
systemic fever often accompanying the illness. The bone changes usually begin before 5 months
of age and resolve before 2 years of age.
Defects in COL1A1 are a cause of Ehlers-Danlos syndrome type 1 (EDS1) [MIM:130000]; also
known as Ehlers-Danlos syndrome gravis. EDS is a connective tissue disorder characterized by
hyperextensible skin, atrophic cutaneous scars due to tissue fragility and joint hyperlaxity. EDS1 is
the severe form of classic Ehlers-Danlos syndrome.
Defects in COL1A1 are the cause of Ehlers-Danlos syndrome type 7A (EDS7A) [MIM:130060];
also known as autosomal dominant Ehlers-Danlos syndrome type VII. EDS is a connective tissue
disorder characterized by hyperextensible skin, atrophic cutaneous scars due to tissue fragility
and joint hyperlaxity. EDS7A is marked by bilateral congenital hip dislocation, hyperlaxity of the
joints, and recurrent partial dislocations.
Defects in COL1A1 are a cause of osteogenesis imperfecta type 1 (OI1) [MIM:166200]. A
dominantly inherited connective tissue disorder characterized by bone fragility and blue sclerae.
Osteogenesis imperfecta type 1 is non-deforming with normal height or mild short stature, and no
dentinogenesis imperfecta.
Defects in COL1A1 are a cause of osteogenesis imperfecta type 2A (OI2A) [MIM:166210]; also
known as osteogenesis imperfecta congenita. A connective tissue disorder characterized by
bone fragility, with many perinatal fractures, severe bowing of long bones, undermineralization,
and death in the perinatal period due to respiratory insufficiency.
Defects in COL1A1 are a cause of osteogenesis imperfecta type 3 (OI3) [MIM:259420]. A
connective tissue disorder characterized by progressively deforming bones, very short stature, a
triangular face, severe scoliosis, grayish sclera, and dentinogenesis imperfecta.
Defects in COL1A1 are a cause of osteogenesis imperfecta type 4 (OI4) [MIM:166220]; also
known as osteogenesis imperfecta with normal sclerae. A connective tissue disorder
characterized by moderately short stature, mild to moderate scoliosis, grayish or white sclera and
dentinogenesis imperfecta.
Genetic variations in COL1A1 are a cause of susceptibility to osteoporosis (OSTEOP)
[MIM:166710]; also known as involutional or senile osteoporosis or postmenopausal
osteoporosis. Osteoporosis is characterized by reduced bone mass, disruption of bone
microarchitecture without alteration in the composition of bone. Osteoporotic bones are more at
risk of fracture.
Note=A chromosomal aberration involving COL1A1 is found in dermatofibrosarcoma
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 protuberans. Translocation t(17;22)(q22;q13) with PDGF.
Sequence similarities Belongs to the fibrillar collagen family.
Contains 1 fibrillar collagen NC1 domain.
Contains 1 VWFC domain.
Post-translational Proline residues at the third position of the tripeptide repeating unit (G-X-Y) are hydroxylated in
modifications some or all of the chains. Proline residues at the second position of the tripeptide repeating unit
(G-X-Y) are hydroxylated in some of the chains.
O-linked glycan consists of a Glc-Gal disaccharide bound to the oxygen atom of a post-
translationally added hydroxyl group.
Cellular localization Secreted > extracellular space > extracellular matrix.

Images

SimpleStep ELISA technology allows the formation of the antibody-

antigen complex in one single step, reducing assay time to 90
minutes. Add samples or standards and antibody mix to wells all at
once, incubate, wash, and add your final substrate. See protocol for
a detailed step-by-step guide.

Other - Human Pro-Collagen I alpha 1 ELISA Kit

(ab210966)

Background-subtracted data values (mean +/- SD) are graphed.

Example of human Pro-Collagen I alpha 1 standard

curve in Sample Diluent NS.

4
 Background-subtracted data values (mean +/- SD) are graphed.

Example of human Pro-Collagen I alpha 1 standard

curve in Sample Diluent 1X Cell Extraction Buffer
PTR.

The concentrations of Pro-Collagen I alpha 1 were measured in

duplicates, interpolated from the Pro-Collagen I alpha 1 standard
curves and corrected for sample dilution. Undiluted samples are as
follows: serum 1%, plasma (citrate) 1%, plasma (EDTA) 1%, and
plasma (heparin) 1%. The interpolated dilution factor corrected
values are plotted (mean +/- SD, n=2). The mean Pro-Collagen I
alpha 1 concentration was determined to be 142.1 ng/mL in serum,
135.9 ng/mL in plasma (citrate), 112.1 ng/mL in plasma (EDTA)
and 102.1 ng/mL in plasma (heparin).

Interpolated concentrations of native Pro-Collagen I

alpha 1 in human serum and plasma samples.

The concentrations of Pro Collagen I alpha 1 were measured in

duplicate and interpolated from the Pro Collagen I alpha 1 standard
curve and corrected for sample dilution. The interpolated dilution
factor corrected values are plotted (mean +/- SD, n=2). The mean
Pro-Collagen I alpha 1 concentration was determined to be 1.62
ng/mL in IMR-90 extract.

Interpolated concentrations of native Pro-Collagen I

alpha 1 in human IMR-90 extract based on a 2
µg/mL extract load.

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 Interpolated dilution factor corrected values are plotted (mean +/-
SD, n=2). The mean Pro-Collagen I alpha 1 concentration was
determined to be 197.3 ng/mL with a range of 113.0 – 417 ng/mL.

Serum from ten individual healthy human female

donors was diluted 1:200 and measured in duplicate.

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