Collagen III alpha 1/COL3A1 Antibody

Immunohistochemistry: Collagen III alpha 1/COL3A1 Antibody [NB600-594] - 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: FFPE. Antigen retrieval: not required. Primary antibody: Anti-collagen type I at 1:500 for 4 degrees Celsius 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.

Western Blot: Collagen III alpha 1/COL3A1 Antibody [NB600-594] - Lane 1: Human Collagen III Load: 100 ng per lane Primary antibody: Collagen III Antibody at 1:1000 o/n at 4C Secondary antibody: DyLight 649 Goat anti-rabbit at 1:20,000 for 30 min at RT Block: incubated with blocking buffer for 30 min at RT Predicted/Observed size: 138 kDa/138 kDa.

Immunocytochemistry/Immunofluorescence: Collagen III alpha 1/COL3A1 Antibody [NB600-594] - Human primary ventricular cardiac fibroblasts were stained with anti-Collagen III antibody. Cells were cultured for 3 days in DMEM with 10% fetal calf serum. ICC/IF image submitted by a verified customer review.

Immunohistochemistry: Collagen III alpha 1/COL3A1 Antibody [NB600-594] - 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 at 99-100C. 20 minutes for antigen retrieval.

Western Blot: Collagen III alpha 1/COL3A1 Antibody [NB600-594] - Immunodetection of ID3, COL1A2 and COL3A1 in human dermal fibroblasts subjected to treatment with or without cANGPTL4 (12 ug/mL) in the presence of TGFB1 (10 ng/mL). B-tubulin, as loading control, was from the same samples. Image collected and cropped by CiteAb from the following publication (www.nature.com/articles/s41598-017-05869-x) licensed under a CC-BY licence.

Immunohistochemistry: Collagen III alpha 1/COL3A1 Antibody [NB600-594] - Collagen I and collagen III accumulate in C57BL/6 mice in response to radiation. Bladder strips from irradiated and control mice were stained for collagen I or for collagen III and imaged using fluorescent microscopy. (a) Representative images (100 x 100 um) of each mouse strain and treatment group are provided for collagen I and collagen III immunofluorescence. Collagen I antibody (NB600-450, RRID:AB_522923) and Collagen III antibody (NB600-594, RRID:AB_10001330). Image collected and cropped by CiteAb from the following publication (onlinelibrary.wiley.com/doi/abs/10.14814/phy2.14377) licensed under a CC-BY licence.

Western Blot: Collagen III alpha 1/COL3A1 Antibody [NB600-594] - Western blot for COL3A1 protein with PVDF protein staining to control for protein loading. Image collected and cropped by CiteAb from the following publication (mdpi.com/2076-3921/6/4/87), licensed under a CC-BY licence.

Immunocytochemistry/Immunofluorescence: Collagen III alpha 1/COL3A1 Antibody [NB600-594] - Collagen III alpha 1 expression in HT-1080 cells. ICC/IF image submitted by a verified customer review.

Immunohistochemistry-Paraffin: Collagen III alpha 1/COL3A1 Antibody [NB600-594] - Human lung tissue.

Immunohistochemistry-Paraffin: Collagen III alpha 1/COL3A1 Antibody [NB600-594] - Human lung tissue

Immunohistochemistry: Collagen III alpha 1/COL3A1 Antibody [NB600-594] - Collagen III (k, positive staining indicated by arrows) immunohistochemistry staining in 13 weeks old male offspring. Left panel = Sham, Middle panel = SE (cigarette smoke exposure), Right panel = SE+MQ (SE with MitoQ supplementation) bar = 50 um. Image collected and cropped by CiteAb from the following publication (mdpi.com/2072-6643/11/7/1669) licensed under a CC-BY licence.

• Reactivity: Hu, Mu, Rt, Bv, Fe, Sh
• Applications: WB, Simple Western, ELISA, ICC/IF, IHC, IHC-P, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated

Collagen III alpha 1/COL3A1 Antibody Summary

• Immunogen: Collagen III alpha 1/COL3A1 from human and bovine placenta (Uniprot: P02461)
• Localization: Extracellular matrix
• Specificity: Typically negligible cross-reactivity against other types of collagens was detected by ELISA against purified standards. Some class-specific anti-collagens may be specific for three-dimensional epitopes which may result in diminished reactivity with denatured collagen or formalin-fixed, paraffin embedded tissues. Non-specific cross-reaction of anti-collagen antibodies with other human serum proteins or non-collagen extracellular matrix proteins is negligible.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: COL3A1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:5000-1:50000
  • Immunocytochemistry/Immunofluorescence 1:10 - 1:500
  • Immunohistochemistry 1:50-1:200
  • Immunohistochemistry-Paraffin 1:50 - 1:200
  • Immunoprecipitation 1:100
  • Simple Western
  • Western Blot 1:1000-1:10000
• Application Notes: Collagen antibodies have been used for indirect trapping ELISA for quantitation of antigen in serum using a standard curve, immunoprecipitation, immunohistochemistry, native (non-denaturing, non-dissociating) PAGE, and western blotting for highly sensitive qualitative analysis.
  
  Simple Western reported by an internal validation. Separated by Size-All, antibody dilution of 1:50. Apparent MW in kDa on Simple Western was 139 kDa; matrix was 12-230 kDa.
• Theoretical MW: 139 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

Use in Sheep reported in scientific literature (PMID:34451177). This antibody reacts with most mammalian Type III collagens and has negligible cross-reactivity with Type I, II, IV, V or VI collagens.
Mouse reactivity reported in multiple pieces of scientific literature. Rat reactivity reported in scientific literature (PMID: 23370982). Feline reactivity reported in scientific literature (PMID: 33091431).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. For extended storage, add an equal volume of glycerol, aliquot and store at -20C or below. Avoid repeated freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Purity: Immunogen affinity purified

Notes

This antibody has been prepared by immunoaffinity chromatography using immobilized antigens followed by extensive cross-adsorption against other collagens, human serum proteins and non-collagen extracellular matrix proteins to remove any unwanted specificities

Store vial at 4C prior to opening. This product is stable at 4C as an undiluted liquid. Dilute only prior to immediate use. For extended storage, mix with an equal volume of glycerol, aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing.

Alternate Names for Collagen III alpha 1/COL3A1 Antibody

• alpha1 (III) collagen
• alpha-1 type III collagen
• COL3A1
• Collagen 3
• collagen alpha-1(III) chain
• Collagen III alpha 1
• collagen, fetal
• collagen, type III, alpha 1
• Collagen-3
• collagenIII
• Collagen-III
• EDS4A
• EDSVASC
• Ehlers-Danlos syndrome type IV, autosomal dominant
• FLJ34534
• PMGEDSV

Background

Collagen III alpha 1, also referred to as collagen type III alpha 1 or COL3A1 for short, was first described in 1971 and is a member of the collagen superfamily and encoded COL3A1 gene (1, 2). In general, collagen III is an extracellular matrix protein that is synthesized as a preprocollagen followed by cleaving of the signal peptide to form the procollagen (1). The human COL3A1 gene is located on chromosome 2q32.2 and collagen III is synthesized as a homotrimer consisting of three identical alpha procollagen chains which are stabilized by disulfide bonds (1,2,3). Each alpha chain is 1466 amino acids (aa) in length with a theoretical molecular weight of 139 kDa for a single alpha chain (1). Structurally, each alpha chain is a left-handed helix which then join together to form a right-handed triple helix (1,2). C-terminal and N-terminal proteinases remove the globular ends of the procollagen to form the type III collagen (1).

Collagen III is a fibrillar collagen that constitutes 5-20% of all collagen in the body (1). It provides structural integrity and is found in many hallow organs and soft connective tissue including the vascular system, skin, lung, uterus, and intestine (1,2). Additionally, collagen III has be found to be associated with type I collagen in the same fibrils (1). Collagen III interacts with signaling integrins to carry out other key functions including cell adhesion, proliferation, and differentiation (1).

Mutations in the COL3A1 gene has been associated with a variety of human diseases, the most well-known being a group of connective tissue disorders termed Ehlers-Danlos Syndromes (1,2,4). Vascular Ehlers-Danlos Syndrome is a specific subtype that is considered the most severe and although the clinical manifestations vary, symptoms include thin skin and fragile blood vessels and can often result in both lung and heart complications (1,4). COL3A1 is also associated with glomerulopathies, or diseases of the glomeruli, which are characterized by an abundance of extracellular matrix (3). Collagenofibrotic glomerulopathy is one specific rare renal disease that is characterized by excessive levels of collagen III (3).

References

1. Kuivaniemi, H., & Tromp, G. (2019). Type III collagen (COL3A1): Gene and protein structure, tissue distribution, and associated diseases. Gene. https://doi.org/10.1016/j.gene.2019.05.003

2. Ricard-Blum S. (2011). The collagen family. Cold Spring Harbor perspectives in biology. https://doi.org/10.1101/cshperspect.a004978

3. Cohen A. H. (2012). Collagen Type III Glomerulopathies. Advances in chronic kidney disease. https://doi.org/10.1053/j.ackd.2012.02.017

4. Olson, S. L., Murray, M. L., & Skeik, N. (2019). A Novel Frameshift COL3A1 Variant in Vascular Ehlers-Danlos Syndrome. Annals of vascular surgery. https://doi.org/10.1016/j.avsg.2019.05.057

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Collagen III alpha 1/COL3A1 Antibody (NB600-594)(28)

Publications using NB600-594

• Willemse J, Verstegen M, Vermeulen A et al. Fast, robust and effective decellularization of whole human livers using mild detergents and pressure controlled perfusion Materials Science and Engineering: C Sep 12 2019 [PMID: 31923991] (IHC-P, IHC, Human)
• Dahl SL, Kypson AP, Lawson JH et al. Readily available tissue-engineered vascular grafts. Sci Transl Med 2011 Feb 2 [PMID: 21289273]
• Mahapatra S Secretion of Cardioprotective Factors from Cardiac Inducing Colonies (Cics) Derived from Germline Pluripotent Stem Cells (Hgpscs) to Combat Myocardial Fibrosis Thesis Jul 13 1905
• Guillaume E, Zacharopoulou M, Reynolds B et al. Additional Value of Second Harmonic Generation Microscopy in the Diagnosis of Feline Collagen Type III Glomerulopathy Journal of Comparative Pathology Oct 1 2021 [PMID: 34686276] (IHC-P, Feline)
• Maity P, Singh K, Krug L Et al. Persistent JunB activation in fibroblasts disrupts stem cell niche interactions enforcing skin aging Cell reports Aug 31 2021 [PMID: 34469740] (ICC/IF, Mouse)
• Oka SI, Byun J, Huang CY, Imai N, Ralda G, Zhai P, Xu X, Kashyap S, Warren JS, Alan Maschek J, Tippetts TS, Tong M, Venkatesh S, Ikeda Y, Mizushima W, Kashihara T, Sadoshima J. Nampt Potentiates Antioxidant Defense in Diabetic Cardiomyopathy Apr 30 2021 [PMID: 33928788] (WB, Mouse)
• Li W, Qi N, Wang S et al. miR-455-5p regulates atrial fibrillation by targeting suppressor of cytokines signaling 3 Journal of physiology and biochemistry Apr 1 2021 [PMID: 33792885]
• Huang L, Zhang Y, Yang J et al. EGR 1 plays a protective role in AMPK inhibitor compound C induced apoptosis through ROS induced ERK activation in skin cancer cellsAnti fibrotic effects and the mechanism of action of miR 29c in silicosis Molecular medicine reports Apr 1 2021 [PMID: 33649785]
• Li H, Wang Y, Liu J, et al. Endothelial Klf2-Foxp1-TGF beta signal mediates the inhibitory effects of simvastatin on maladaptive cardiac remodeling Theranostics Jan 1 2021 [PMID: 33408770] (WB)
• Paul K, Darzi S, Del Borgo M et al. Vaginal delivery of tissue engineered endometrial mesenchymal stem/stromal cells in an aloe vera-alginate hydrogel alleviates maternal simulated birth injury Applied Materials Today Mar 1 2021 (ICC/IF, Human)
• Oikawa K, Teixeira LBC, Keikhosravi A et al. Microstructure and resident cell-types of the feline optic nerve head resemble that of humans Exp Eye Res Oct 19 2020 [PMID: 33091431] (ICC/IF, Feline)
• Birchmeier C, Akalin A, Spuler S et al. Single-nucleus transcriptomics reveals functional compartmentalization in syncytial skeletal muscle cells bioRxiv Jan 1 2020 (IHC, Mouse)
• Zwaans Bmm, Wegner Ka, Bartolone Sn et Al. Radiation cystitis modeling: A comparative study of bladder fibrosis radio-sensitivity in C57BL/6, C3H, and BALB/c mice Physiol Rep Feb 1 2020 [PMID: 32109348] (ICC/IF, Mouse)
• Willemse J, Verstegen M, Vermeulen A et al. Fast, robust and effective decellularization of whole human livers using mild detergents and pressure controlled perfusion Materials Science and Engineering: C Sep 12 2019 (IHC-P, IHC, Human)
• Antonova LV, Krivkina EO, Sevostianova VV Et al. Tissue-Engineered Carotid Artery Interposition Grafts Demonstrate High Primary Patency and Promote Vascular Tissue Regeneration in the Ovine Model Polymers Aug 8 2021 [PMID: 34451177] (IHC, Sheep)
• Sales VL, Engelmayr GC Jr, Johnson JA Jr et al. Protein precoating of elastomeric tissue-engineering scaffolds increased cellularity, enhanced extracellular matrix protein production, and differentially regulated the phenotypes of circulating endothelial progenitor cells Circulation [PMID: 17846326]
  
  Details:
  Used the Biotin form of this antibody.
• Willemse J, Verstegen M, Vermeulen A et al. Fast, robust and effective decellularization of whole human livers using mild detergents and pressure controlled perfusion Materials Science and Engineering: C Sep 1 2019 (IHC, Human)
• Guo Xiangjiang, Fereydooni Arash, Isaji Toshihiko et al. Inhibition of the Akt1-mTORC1 Axis Alters Venous Remodeling to Improve Arteriovenous Fistula Patency. Scientific Reports 2019 Jul 30 [PMID: 31363142] (IHC-P, Mouse)
• Yang J, Wang B, Li N et al. Salvia miltiorrhiza and Carthamus tinctorius Extract Prevents Cardiac Fibrosis and Dysfunction after Myocardial Infarction by Epigenetically Inhibiting Smad3 Expression Evid Based Complement Alternat Med. May 30 2019 [PMID: 31275414] (WB, Mouse)
• Wang X, Pan J, Liu H et al. AIM2 gene silencing attenuates diabetic cardiomyopathy in type 2 diabetic rat model Life Sci. Feb 18 2019 [PMID: 30790610] (IHC-P, WB, Rat)
• Zhang M, Zhu P, Wang Y et al. Bilateral sympathetic stellate ganglionectomy attenuates myocardial remodelling and fibrosis in a rat model of chronic volume overload. J. Cell. Mol. Med. Nov 8 2018 [PMID: 30411499] (WB, Rat)
• Angel PM, Comte-Walters S, Ball LE et al. Mapping Extracellular Matrix Proteins in Formalin-Fixed, Paraffin-embedded Tissues by MALDI Imaging Mass Spectrometry J. Proteome Res. 2017 Nov 21 [PMID: 29161047] (IHC-P)
• Schreckenberg R, Horn AM, da Costa Rebelo RM et al. Effects of 6-months' Exercise on Cardiac Function, Structure and Metabolism in Female Hypertensive Rats-The Decisive Role of Lysyl Oxidase and Collagen III Front Physiol 2017 Aug 21 [PMID: 28824452] (WB, Rat)
  
  Details:
  Collagen III antibody was used to look at the effect of running training on the expression of COL III in rats that were either sedentary or subjected to running training
• Teo Z, Chan JSK, Chong HC et al. Angiopoietin-like 4 induces a B-catenin-mediated upregulation of ID3 in fibroblasts to reduce scar collagen expression Sci Rep 2017 Jul 24 [PMID: 28740178] (WB, Human)
• Lee HJ, Kim YS, Ok JH et al. Effect of a single subacromial prednisolone injection in acute rotator cuff tears in a rat model. Knee Surg Sports Traumatol Arthrosc 2013 Jan 31 [PMID: 23370982] (IHC, Rat)
• Mak KM, Chu E, Lau KH, Kwong AJ. Liver fibrosis in elderly cadavers: localization of collagen types I, III, and IV, alpha-smooth muscle actin, and elastic fibers. The Anatomical Record, 2012; vol 295, pages 1159-1167. 2012 Jul [PMID: 22644959]
• Mak KM, Kwong AJ, Chu E et al. Hepatic steatosis, fibrosis, and cancer in elderly cadavers. The Anatomical Record, 2012; vol 295, pages 40-50. 2012 Jan [PMID: 22139908]
• Hechler B, Gachet C Comparison of two murine models of thrombosis induced by atherosclerotic plaque injury. Thromb Haemost 2011 May [PMID: 21479341] (IHC, Mouse)

Reviews for Collagen III alpha 1/COL3A1 Antibody (NB600-594) (7)

Average Rating: 4.7

(Based on 7 reviews)

We have 7 reviews tested in 4 species: Human, Mouse, Rat, Feline.

reviewed by: Shinichi Oka WB Mouse 04/07/2021

Summary

• Application: Western Blot
• Sample Tested: Heart lysates
• Species: Mouse
• Lot: 42255

Comments

• Comments: Mouse heart lysate was prepared after 3 months of normal diet (ND) and high fat diet (HFD) consumption. Western blot was performed with anti-Col3a1 antibody.

reviewed by: Verified Customer WB Human 06/07/2018

Summary

• Application: Western Blot
• Sample Tested: whole cell lysate
• Species: Human
• Lot: 37149

reviewed by: Verified Customer IHC-Fr Feline 02/18/2018

Summary

• Application: Immunohistochemistry-Frozen
• Sample Tested: Connective tissue
• Species: Feline
• Lot: 39661

reviewed by: wang xuyang IHC-P Rat 12/20/2017

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: H9c2 whole cell lysate
• Species: Rat
• Lot: lot37797

reviewed by: Verified Customer IHC-P Rat 07/20/2016

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: rat colon
• Species: Rat
• Lot: 33577

reviewed by: Jessica Ochoa IF Human 04/02/2015

Summary

• Application: Immunofluorescence
• Sample Tested: HT-1080 (Fibrosarcoma)
• Species: Human

Blocking

• Blocking Details: 5% BSA (prepared in 1X PBS) for 20 minutes at room temperature

Primary Anitbody

• Dilution Ratio: 1:1000, 1 hour, room temperature

Secondary Antibody

• Secondary Description: Goat Anti-Rabbit IgG, Alexa Fluor 594
• Secondary Manufacturer Cat#: A11012
• Secondary Concentration: 1:500 dilution

Details

• Detection Notes: HT-1080 (positive control), Human Astrocytes (negative control)
• Fixation Details: 4% PFA fix for 20 minutes at room temperature
• Wash Description: 1X PBS, 5 minutes, 3 washes

reviewed by: Verified Customer IF Human 10/04/2014

Summary

• Application: Immunofluorescence
• Sample Tested: human primary ventricular cardiac fibroblasts (from ScienCell)
• Species: Human
• Lot: 30240

Blocking

• Blocking Details: 1 mg/ml BSA in PBS, 30 min, room temperature

Primary Anitbody

• Dilution Ratio: 1:100 in BSA/PBS overnight at 4C

Secondary Antibody

• Secondary Description: Goat anti rabbit Alexa488
• Secondary Manufacturer Cat#: A-21222
• Secondary Concentration: 1:200

Details

• Detection Notes: confocal microscopy, Zeiss LSM710
• Fixation Details: fixation with 3% PFA in PBS for 15 min at room temperature
• Wash Description: washes with PBS, usually 3 times for 5 minutes

FAQs for Collagen III alpha 1/COL3A1 Antibody (NB600-594). (Showing 1 - 3 of 3 FAQs).

1. May I know if you carry the un-conjugated version of NB120-6580 (biotinylated anti-Collagen Type iII)?
   • I would recommend this catalog number for an unconjugated anti-Collagen Type III antibody: NB600-594.
2. What is the dilution factor for IHC for starting? Because the range of dilution factor on the datasheet is so broad, I am confused which dilution factor is suitable for starting.
   • I can recommend the following dilution to start. Keep in mind that these will all vary greatly by sample type, so these are just suggestions. You may need to use more or less antibody. For catalog number NB600-594, the dilution range is 1:50-1:200.
3. I am wondering if you could please confirm for me whether this product Collagen III antibody - catalogue # NB600-594 is able to be used under denaturing conditions with Western blots? Or whether it only binds to the collagen in its natural (non-denatured)
   • The Western blot image is not generated from native collagen - reducing conditions were used to obtain the image. The intact native molecule is too large to be run on standard 4-20% gels so the sample was run under reducing conditions. The image shows the sub-unit strand(s), not the intact collagen that consists of 3 subunit strands. These are not considered optimal or specific binding conditions for the anti-collagen antibodies, however the decision was made to run and post this on the website as an example.

Bioinformatics

• Gene Symbol: COL3A1