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

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. Image from verified customer review.

Immunohistochemistry: Collagen III alpha 1/COL3A1 Antibody [NB600-594] - Analysis of Biotin conjugate of NB600-594. 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 a

Immunocytochemistry/Immunofluorescence: Collagen III alpha 1/COL3A1 Antibody [NB600-594] - Collagen III alpha 1 expression in HT-1080 cells. Image from verified customer review.

Immunohistochemistry-Paraffin: 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: formalin fixed paraffin embedded. 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.

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] - 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.

Immunohistochemistry: Collagen III alpha 1/COL3A1 Antibody [NB600-594] - Analysis of FITC conjugate of 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 s

• Reactivity: Hu, Mu, Rt, Bv, Ma
• Applications: WB, ELISA, ICC/IF, IHC, IHC-P, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

Collagen III alpha 1/COL3A1 Antibody Summary

• Immunogen: Collagen Type III from human and bovine placenta.
• Localization: Extracellular matrix
• Specificity: Typically less than 1% 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. This antibody reacts with most mammalian Type III collagens and has 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.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: COL3A1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:1000-1:10000
  • 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
• Application Notes: Greatly diminished reactivity and selectivity of these antibodies will result if denaturing and reducing conditions are used for in immunoblotting.
  ICC/IF data from customer review.

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 21479341). Rat reactivity reported in scientific literature (PMID: 23370982).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 0.125M Sodium Borate, 75mM NaCl and 5mM EDTA, pH 8.0
• Preservative: 0.01% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for Collagen III alpha 1/COL3A1 Antibody

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

Background

These antibodies are well suited to detect extracellular matrix proteins in normal as well as disease state tissues. Disruption of tissue organization is the hallmark of neoplasia. Malignant lesions can be distinguished from benign by examining the breakdown of basement membranes and loss of 3-dimensional architecture. Malignant cells are presumed to use matrix metalloproteases to degrade barriers created by the extracellular matrix which then allows metastasis to occur. Collagenases, stomelysins and gelatinases can collectively degrade all of the various components of the extracellular matrix, including fibrillar and non-fibrillar collagens and basement membrane glycoproteins.

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)(8)

Publications using NB600-594

• 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)
• Dahl SL, Kypson AP, Lawson JH et al. Readily available tissue-engineered vascular grafts. Sci Transl Med 2011 Feb 2 [PMID: 21289273]
• 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.
• 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
  
  Details:
  This publication used the Biotin conjugated form of this antibody (Cat# NB120-6580).

Reviews for Collagen III alpha 1/COL3A1 Antibody (NB600-594) (3)

Average Rating: 4.7

(Based on 3 reviews)

We have 3 reviews tested in 2 species: Human, Rat.

We have 3 reviews tested in 2 application: IHC-P, IF.

reviewed by: Anonymous IHC-P Rat 07/20/2016

Summary

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

reviewed by: Jessica Prieto 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: Anonymous 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
• Entrez:
  • 1281(Human)
  • 12825(Mouse)
  • 84032(Rat)