Overview

  • Product nameAnti-Collagen III antibodySee all Collagen III primary antibodies ...
  • Description
    Rabbit polyclonal to Collagen III
  • SpecificityThis type specific collagen antibody only recognizes 3D epitopes. Negligible cross-reactivity with Type I, II, IV, V or VI collagens. Non-specific cross reaction of anti-collagen antibodies with other human serum proteins or non-collagen extracellular matrix proteins is negligible
  • Tested applicationsIHC-P, ICC/IF, ICC, ELISA, IHC-Fr, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
    Predicted to work with: all Mammals
  • Immunogen

    Full length native protein (purified) corresponding to Collagen III aa 1-1466.
    Database link: P02461

  • Positive control
    • Human skin. Human testicle tissue.
  • General notes

    At least 11 genetically distinct gene products are collectively referred to as 'collagen types' or other proteins and proteoglycans of the extracellular matrix. In humans, collagens are composed of about 20 unique protein chains which under go various types of post-translational modifications and are ultimately assembled into a triple helix. This results in great diversity between collagen types. Collagens are highly conserved throughout evolution and are characterized by an uninterrupted "Glycine-X-Y" triplet repeat that is a necessary part of the triple helical structure. For these reasons it is often extremely difficult to generate antibodies with specificities to collagens. The development of type specific antibodies is dependent on NON-DENATURED three-dimensional epitopes. This preparation results in a native conformation of the protein.

Properties

Applications

Our Abpromise guarantee covers the use of ab7778 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
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent dilution. PubMed: 18385800
ICC 1/100. PubMed: 19036760
ELISA 1/1000 - 1/8000. This product was assayed against 1.0 ug of Collagen III in a direct ELISA using Peroxidase conjugated Goat anti-Rabbit and ABTS (2,2'-azino-bis-[3- ethylbenthiazoline-6-sulfonic acid]) as a substrate for 30 minutes at room temperature. This product can also be used with Biotin Conjugated Anti-Collagen III (ab6580) in a sandwich ELISA.
IHC-Fr 1/50 - 1/200.
IP 1/5000 - 1/10000.
WB 1/5000 - 1/10000. Predicted molecular weight: 138 kDa.

Target

  • FunctionCollagen type III occurs in most soft connective tissues along with type I collagen.
  • Involvement in diseaseDefects in COL3A1 are a cause of Ehlers-Danlos syndrome type 3 (EDS3) [MIM:130020]; also known as benign hypermobility syndrome. EDS is a connective tissue disorder characterized by hyperextensible skin, atrophic cutaneous scars due to tissue fragility and joint hyperlaxity. EDS3 is a form of Ehlers-Danlos syndrome characterized by marked joint hyperextensibility without skeletal deformity.
    Defects in COL3A1 are the cause of Ehlers-Danlos syndrome type 4 (EDS4) [MIM:130050]. EDS is a connective tissue disorder characterized by hyperextensible skin, atrophic cutaneous scars due to tissue fragility and joint hyperlaxity. EDS4 is the most severe form of the disease. It is characterized by the joint and dermal manifestations as in other forms of the syndrome, characteristic facial features (acrogeria) in most patients, and by proneness to spontaneous rupture of bowel and large arteries. The vascular complications may affect all anatomical areas.
    Defects in COL3A1 are a cause of susceptibility to aortic aneurysm abdominal (AAA) [MIM:100070]. AAA is a common multifactorial disorder characterized by permanent dilation of the abdominal aorta, usually due to degenerative changes in the aortic wall. Histologically, AAA is characterized by signs of chronic inflammation, destructive remodeling of the extracellular matrix, and depletion of vascular smooth muscle cells.
  • Sequence similaritiesBelongs 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.
    O-linked glycan consists of a Glc-Gal disaccharide bound to the oxygen atom of a post-translationally added hydroxyl group.
  • Cellular localizationSecreted > extracellular space > extracellular matrix.
  • Information by UniProt
  • Database links
  • Alternative names
    • CO3A1_HUMAN antibody
    • COL 3A1 antibody
    • COL3A1 antibody
    • Collagen alpha 1(III) chain antibody
    • Collagen alpha-1(III) chain antibody
    • Collagen III alpha 1 chain precursor antibody
    • Collagen III alpha 1 polypeptide antibody
    • Collagen type III alpha 1 (Ehlers Danlos syndrome type IV autosomal dominant) antibody
    • Collagen type III alpha 1 antibody
    • Collagen type III alpha antibody
    • EDS4A antibody
    • Ehlers Danlos syndrome type IV, autosomal dominant antibody
    • Fetal collagen antibody
    • Type III collagen antibody
    see all

Anti-Collagen III antibody images

  • Ab53088 staining Human skin (ab30166). Staining is localised to the extracellular matrix.
    Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the DAKO 3-in-1 antigen retrieval buffer citrate pH 6.0 (ab64214) in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

  • ab7778 at 1:400 (45 min RT) showed strong staining in FFPE sections of human skin(left, dermis) with moderate to strong red staining and testis (right) where strong staining was observed within connective tissue between seminiferous tubules. The antibody showed strong extracellular staining within connective tissues across many organs with minimal background staining. Slides were steamed in 0.01 M sodium citrate buffer, pH 6.0 (ab64214) at 99-100°C - 20 minutes for antigen retrieval.

  • ab7778 staining Collagen III in human testicle tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    Tissue was fixed with paraformaldehyde and a heat mediated antigen retrieval step was performed using TE buffer pH 9.0. Samples were then incubated with ab7778 at a 1/200 dilution for 30 minutes at 20°C. The secondary used was an undiluted, HRP-conjugated goat anti-rabbit polyclonal.

    See Abreview

  • Immunohistochemistry of ab7778. Tissue: right lobe of the liver section. A:Central Vein (CV) fibrosis, B: Non-fibrotic CV, C: Perisinusodial fibrosis, D: Non-fibrotic area, E: Protat tract fibrosis, F: Septal fibrosis (arrow). Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: Anti-collagen type I at 1:500 for 4°C for 24hr. Secondary antibody: Peroxidase biotin-streptavidin rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: Anti-collagen type III is intra and extracellular. Staining: 3.3’-diaminobenzidine tetrahydrochloride was used as the chromogen. Nuclei were counterstained purple with hematoxylin.
  • ab7778 staining Collagen III in Human saphenous vein-derived myofibroblasts by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton-X100. Prior to staining, cells were washed and blocked in a 2% bovine serum albumin, 0.1% saponin (w=v) solution in PBS, and subsequently incubated for 1 hour with the primary antibody in PBS. Nuclei are stained with Hoechst.
  • Anti-Collagen III antibody (ab7778) at 1/1000 dilution + BME

    Predicted band size : 138 kDa
    Western Blot produced under denaturing and reducing conditions.

References for Anti-Collagen III antibody (ab7778)

This product has been referenced in:
  • Kach J  et al. Antifibrotic effects of noscapine through activation of prostaglandin E2 receptors and protein kinase A. J Biol Chem 289:7505-13 (2014). IHC (PFA fixed) ; Human . Read more (PubMed: 24492608) »
  • Takase HM  et al. FGF7 is a functional niche signal required for stimulation of adult liver progenitor cells that support liver regeneration. Genes Dev 27:169-81 (2013). Read more (PubMed: 23322300) »

See all 16 Publications for this product

Product Wall

Thank you for contacting us.

I can confirm that any general western blotting protocol would work with this antibody. Please check the following conditions;

- the conditions should be native and non-reduced.
- we normally recommend...

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Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Sample Mouse Cell (Hepatocyte)
Specification Hepatocyte
Permeabilization No
Fixative Paraformaldehyde
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Submitted Feb 17 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Liver)
Specification Liver
Fixative Paraformaldehyde
Permeabilization No
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Apr 26 2013

We have issued a free of charge replacement.

Thank you for contacting us.

The size of native protein is predicted as 138 kDa (http://www.uniprot.org/uniprot/P02461). The observed size will be slightly different due to post translational changes to proteins. Please check literature what o...

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Unfortunately we don't have free of charge controls available for me to send you. However as a positive control I'd recommend http://www.abcam.com/placenta-human-tissue-lysate-adult-normal-tissue-ab29745.html. Collagen III purified from human...

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Thanks for your enquiry.

It is recommended to run non denaturing gels for separation and blotting of Collagen to detect them specifically, as all collagens have a similar amino acid structure of Gly-X-Y. However I have seen many instances whe...

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Hello! I have attached the image for the ˜129 kD band expected for Collagen I. The laboratory has indicated that the sample preparation does account for some change in MW when using Western Blotting for Collagens. The legend for the image is below. Ple...

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Yes, the sample was reduced with beta mercaptoethanol. Please let me know if you have any other questions.

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

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