Quantitative Analysis of Extracellular Matrix Proteins

Each section was incubated overnight at 4 °C with primary antibodies targeting Type I procollagen (ab64409, rat mAb, Abcam, Cambridge, UK, cross-reacts with human), Type I collagen (600–401-103, rabbit pAb, Rockland, Limerick, PA, cross-reacts with bovine and human), Type III collagen (ab6310, FH-7A, mouse mAb, Abcam, Cambridge, UK, cross-reacts with rat and human), Type V collagen (AM10159PU-N, V13F6, mouse mAb, Acris, San Diego, CA, cross-reacts with human), keratin-14 (20R-CP002, guinea pig pAb, Fitzgerald Industries International, Acton, MA) and PDGFRbeta (MAB1263, mouse mAb, R&D Systems, Minneapolis, MN). Primary antibodies were detected using Alexa488- or Alexa594-conjugated secondary antibody (Thermo Fisher Scientific). Sections were examined with an Olympus BX51 microscope (Olympus, Tokyo, Japan) and images were captured with a DP72 controller digital camera (Olympus). The staining intensity in 6 randomly selected microscopic fields was quantified by using WINROOF2015 image-analyzing software (Mitani, Fukui, Japan, https://www.mitani-visual.jp/products/image_analys_ismeasurement/winroof/), as described previously^{10 (link)}.

Free full text: Click here

Iriyama S., Ogura Y., Nishikawa S., Hosoi J, & Amano S. (2022). Regeneration of collagen fibrils at the papillary dermis by reconstructing basement membrane at the dermal–epidermal junction. Scientific Reports, 12, 795.

Publication 2022

ab64409 ab6310 FH-7A MAB1263 Alexa594-conjugated secondary antibody BX51 microscope DP72 controller digital camera

Alexa594 Antibodies Antibody Bovine Camera Guinea pig Human Keratin 14 Microscopic Mouse Pdgfrbeta Rabbit Type i collagen Type i procollagen Type iii collagen Type v collagen

Top 5 similar protocols

Variable analysis

independent variables

Incubation of sections overnight at 4 °C with primary antibodies targeting Type I procollagen, Type I collagen, Type III collagen, Type V collagen, keratin-14, and PDGFRbeta

dependent variables

Staining intensity of the primary antibodies in 6 randomly selected microscopic fields

control variables

Incubation temperature of 4 °C
Use of specific primary antibodies that cross-react with the target species (human, bovine, rat)

controls

No positive or negative controls were explicitly mentioned in the protocol.

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!