Overview

  • Product name
    Anti-Collagen I antibody [COL-1]
    See all Collagen I primary antibodies
  • Description
    Mouse monoclonal [COL-1] to Collagen I
  • Host species
    Mouse
  • Specificity
    ab90395 will not cross react with collagen 2-11, or thermally denatured collagen. The antibody is reactive with the native (non-denaturing, helical) form of collagen type I and not reactive with hen tested on thermally denatured molecules. Use native (non-denaturing) conditions.
  • Tested applications
    Suitable for: Dot blot, IHC-Glut, IHC-R, IP, Indirect ELISA, WB, ELISA, IHC-Fr, ICC/IF, Electron Microscopy, IHC-FoFrmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Rat, Rabbit, Cow, Human, Pig, Deer
  • Immunogen

    Full length native protein (purified) corresponding to Cow Collagen I.
    Database link: P02453

  • Positive control
    • Connective tissue fibres. Pig skin tissue.
  • General notes

    IHC-P application: We have received positive as well as negative customer feedback for IHC-P. The antibody is not batch-tested in this application, thus additional troubleshooting might be needed to obtain satisfactory results.

Properties

Applications

Our Abpromise guarantee covers the use of ab90395 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Dot blot Use at an assay dependent concentration.

Assay dependent

IHC-Glut Use at an assay dependent concentration.
IHC-R Use at an assay dependent concentration.
IP 1/50.
Indirect ELISA Use at an assay dependent concentration.
WB 1/500 - 1/3000. Use under non reducing condition. Predicted molecular weight: 139 kDa.

The antibody is reactive with the native (non-denaturing, helical) form of collagen type I and not reactive when tested on thermally denatured molecules. Use native (non-denaturing) conditions, as the antibody does not recognise denatured protein.

ELISA Use at an assay dependent concentration.

Assay dependent

IHC-Fr 1/2000.
ICC/IF 1/2000.
Electron Microscopy 1/100 - 1/1000. PubMed: 17016762for Gold labeling
IHC-FoFr 1/100. PubMed: 17016762Fix in Zamboni's solution (2% paraformaldehyde, 0.2% picric acid in phosphate-buffered saline (PBS), pH 7.6) for 2 h at 4C, store in 20% sucrose in 0.5 mM PBS at 4C.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • 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 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
      modifications
      Proline residues at the third position of the tripeptide repeating unit (G-X-Y) are hydroxylated in 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.
    • Information by UniProt
    • Database links
    • Alternative names
      • Alpha 1 type I collagen antibody
      • Alpha 2 type I collagen antibody
      • alpha 2 type I procollagen antibody
      • alpha 2(I) procollagen antibody
      • alpha 2(I)-collagen antibody
      • Alpha-1 type I collagen antibody
      • alpha1(I) procollagen antibody
      • CO1A1_HUMAN antibody
      • COL1A1 antibody
      • COL1A2 antibody
      • collagen alpha 1 chain type I antibody
      • Collagen alpha-1(I) chain antibody
      • collagen alpha-1(I) chain preproprotein antibody
      • Collagen I alpha 1 polypeptide antibody
      • Collagen I alpha 2 polypeptide antibody
      • collagen of skin, tendon and bone, alpha-1 chain antibody
      • collagen of skin, tendon and bone, alpha-2 chain antibody
      • Collagen type I alpha 1 antibody
      • Collagen type I alpha 2 antibody
      • EDSC antibody
      • OI1 antibody
      • OI2 antibody
      • OI3 antibody
      • OI4 antibody
      • pro-alpha-1 collagen type 1 antibody
      • type I proalpha 1 antibody
      • type I procollagen alpha 1 chain antibody
      • Type I procollagen antibody
      see all

    Images

    • All lanes : Anti-Collagen I antibody [COL-1] (ab90395) at 1/1000 dilution

      Lane 1 : Recombinant Human Collagen I at 3 µg
      Lane 2 : Pig skin whole cell lysate extracted in Laemmli buffer at 20 µg

      Secondary
      All lanes : HRP-conjugated Goat anti-mouse monoclonal IgG at 1/3000 dilution

      Developed using the ECL technique.

      Performed under non-reducing conditions.

      Predicted band size: 139 kDa
      Observed band size: 130,140 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 250 kDa (possible dimer), 400 kDa (possible multimer)


      Exposure time: 30 seconds


      Blocked with 5% Milk for 1 hour at 20°C.

      See Abreview

    • ab90395 staining Collagen I in Pig Ureter tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 5% BSA for 30 minutes at 20°C. Samples were incubated with primary antibody (1/2000) for 4 hours at 2°C. A Cy2®-conjugated Goat anti-mouse polyclonal (1/800) was used as the secondary antibody.

      See Abreview

    • ab90395 staining Collagen I in Pig skin tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with formaldehyde and blocked with 5% serum for 1 hour at 20°C. Samples were incubated with primary antibody (1/500 in PBS, Tween 0.01% + Donkey serum 1%) for 16 hours at 4°C. A Cy2® -conjugated Donkey anti-mouse IgG polyclonal (1/100) was used as the secondary antibody.

      See Abreview

    References

    This product has been referenced in:
    • Cunniffe GM  et al. Tissue-specific extracellular matrix scaffolds for the regeneration of spatially complex musculoskeletal tissues. Biomaterials 188:63-73 (2019). Read more (PubMed: 30321864) »
    • Critchley S  et al. Regeneration of Osteochondral Defects Using Developmentally Inspired Cartilaginous Templates. Tissue Eng Part A N/A:N/A (2018). Read more (PubMed: 30358516) »
    See all 85 Publications for this product

    Customer reviews and Q&As

    1-10 of 19 Abreviews or Q&A

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Sheep Tissue sections (Osteochondral section of the knee joint patella)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Sodium citrate pH6, O/N, 60 °C + proteinase K, 15 min, 37 °C
    Permeabilization
    No
    Specification
    Osteochondral section of the knee joint patella
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Fixative
    Paraformaldehyde

    Ms. Amra Secerovic

    Verified customer

    Submitted Dec 07 2018

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Dermal Fibroblast)
    Permeabilization
    No
    Specification
    Dermal Fibroblast
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
    Fixative
    Formaldehyde

    Adrian Djalali Cuevas

    Verified customer

    Submitted Sep 22 2017

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (muscle)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate buffer
    Specification
    muscle
    Blocking step
    (agent) for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Mar 16 2016

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 0.05% typsin
    Sample
    Human Tissue sections (human whole fat tissue)
    Specification
    human whole fat tissue
    Permeabilization
    No
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Mar 13 2015

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: sodium citrate buffer at pH=6.05
    Sample
    Sheep Tissue sections (tendon)
    Specification
    tendon
    Permeabilization
    No
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Dec 13 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Pig Tissue sections (Ureter from Minipig)
    Specification
    Ureter from Minipig
    Fixative
    Paraformaldehyde
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C

    Ms. Beate Thode

    Verified customer

    Submitted Feb 20 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (Ureter)
    Specification
    Ureter
    Fixative
    Paraformaldehyde
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C

    Ms. Beate Thode

    Verified customer

    Submitted Feb 19 2013

    Question
    Answer



    ab90395 is in form of ascites. The concentration is not determined.

    Read More

    Answer

    Thank you for contacting us.

    I have looked at the datasheet and indeed the information is confusing.

    Reducing and non-reducing conditions refers to whether or not there are reagents present that disrupt disulfide bonds between cysteine residues in the protein. Antibody molecules have many disulfide linkages, which are critical to maintaining their structure. When reagents that disrupt disulfide bonds, such as dithiothreitol or mercaptoethanol, are present in the solution, antibody molecules lose their active structure. When these agents are present it is referred to as "reducing" conditions.

    Denaturing conditions are where hydrogen bonds within the protein are broken and therefore the protein's 3D structure unfolds.

    We recommend using non reducing conditions and native (non-denaturing) conditions. This is because the antibody recognises the native form of the protein and therefore we would expect you to get the best results by using non-reducing, non-denaturing conditions. This would involve not using b-mecaptoethanol, DTT, SDS or heating. We would also recommend the use of protease inhibitors to reduce the degradation of the protein.

    The reason we have conflicting information on our datasheets is because we have had some customers who have used different running conditions and have got positive results which they have submitted to us via Abreviews and we publish all these results. This explains why we have a WB image which uses laemmli buffer on our datasheet. This image is from one of our customers who got positive results using these conditions. However, we recommend using non-reducing and non-denaturing conditions for optimal results.

    I am attaching a word document which outlines which buffers should be used.

    Read More

    Answer

    >The AMELX protein ab139212 can be diluted in any buffer. However, all of our products are intended for research use only and are not recommended for in vitro studies. This protein contains DTT, glycerol, and Tris HCL in its buffer, and it is not a sterile product, so we do not suggest it for use in a live animal study.

    Ab13420 and ab90395 are mouse monoclonal antibodies, so they can be detected with a detection kit like ab94743-

    https://www.abcam.com/EXPOSE-Mouse-Specific-AP-red-Detection-IHC-Kit-ab94743.html <

    Read More

    1-10 of 19 Abreviews or Q&A

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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