Eleven primary antibodies, including anti-OSX, anti-COL1A1, anti-OC, anti-MMP-1, anti-MMP-3, andanti-MMP-13, were purchased from Abcam (Cambridge, UK). After culturing for 48 h, the isolated chondrocytes were lysed in radioimmunoprecipitation assay (RIPA) lysis buffer. The total protein concentration was determined using a bicinchoninic acid (BCA) kit (Pierce, Rockford, IL, USA). Equal amounts of protein were isolated by sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride membranes (Bio-Rad, Hercules, CA). The membranes were blocked with 5% skim milk in Tris-buffered saline Tween (TBST) for 1 h, and then incubated with primary antibody anti-OSX (1:1,000, ab22552, Abcam), anti-COL1A1 (1:1,000, ab34710, Abcam), anti-OC (1:500, ab93876, Abcam), anti-MMP-1 (1:1,000, ab137332, Abcam), anti-MMP-3 (1:500, ab53015, Abcam), and anti-MMP-13 (1:3,000, ab39012, Abcam) at 4 °C overnight. The membranes were then washed and subjected to goat anti-rabbit immunoglobulin (Ig)G horseradish peroxidase (HRP)-conjugated secondary antibodies. The blots were detected using an enhanced chemiluminescent (ECL) detection kit (Pierce, Rockford, IL, USA). Actin was used as a loading control. The band densities were determined and analyzed with an automatic digital gel image analysis system Bio-Rad CFX-96 (Bio-Rad, CA, USA).
Anti oc
Manufactured by Abcam
Sourced in United Kingdom, United States
Anti-OC is a laboratory product designed for the detection and quantification of the OC protein. It is a tool used in research applications to study the OC protein and its role in biological processes. The core function of Anti-OC is to provide a reliable and accurate method for the analysis of OC protein levels.
Automatically generated - may contain errors
Lab products found in correlation
Ab39012, by Abcam (1 mentions)
Ab137332, by Abcam (1 mentions)
Ab53015, by Abcam (1 mentions)
Anti osx, by Abcam (1 mentions)
Ab34710, by Abcam (1 mentions)
Ab93876, by Abcam (1 mentions)
Ab22552, by Abcam (1 mentions)
Anti col1a1, by Abcam (1 mentions)
Anti mmp1, by Abcam (1 mentions)
Anti mmp 3, by Abcam (1 mentions)
Cfx96, by Bio-Rad (1 mentions)
Polyvinylidene difluoride membrane, by Bio-Rad (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
β actin, by Abcam (1 mentions)
Anti collagen type 1, by Abcam (1 mentions)
Anti bsp, by Abcam (1 mentions)
Citrate buffer, by Merck Group (1 mentions)
Te2000 microscope, by Nikon (1 mentions)
Anti cd31, by Abcam (1 mentions)
Anti runx2, by Abcam (1 mentions)
4 protocols using anti oc
1
Chondrocyte Protein Expression Profiling
2
Protein Expression Analysis in Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After 3 days of culture, cells were lysed using Nonidet-P40 (NP40) buffer (Thermo), and a BCA protein assay kit was used to assess for the concentrations of the various proteins. Sodium dodecyl sulfate (SDS)–polyacrylamide gel electrophoresis was used to segregate the cell lysates (40 μg protein), which were then transferred onto polyvinylidene difluoride (PVDF) membranes (Merck Millipore, Burlington, MA, USA). These were placed in a solution containing 2% bovine serum albumin (BSA) and Tris-buffered saline with Tween 20 (TBST) for 1 h before they were exposed to primary anti-bone morphogenetic protein receptor type II (BMP2R), anti-phospho SMAD1/5/8, anti-phospho extracellular signal-regulated kinase 1/2 (p-ERK 1/2), anti-extracellular signal-regulated kinase 1/2 (ERK 1/2), anti-ALP, anti-OC, and β-actin (Abcam, Cambridge, MA, USA) for 2 h for further immunoblotting. Thereafter, horseradish peroxidase (HRP)-conjugated secondary antibodies were incubated with the samples for 1 h to enable chemiluminescence visualization. For this study, protein expression levels were normalized to β-actin. All experiments were performed in triplicate.
3
Immunohistochemical Analysis of Bone Regeneration
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The central lateral sections of each group sample were probed with collagen type I, bone sialoprotein (BSP), osteocalcin (OC), and angiogenic marker CD31. Briefly, the samples were retrieved with 1 × citrate buffer (Sigma-Aldrich, St. Louis, MO) for 30 min at 95–100 °C. Followed which, all sets of groups were incubated with primary anti-BSP (1:500), anti-collagen type I (1:250), anti-OC (1:100), and anti-CD31 (1:50) antibodies (Abcam, Cambridge, UK) separately in a humidity chamber at 4 °C for overnight, and were subsequently marked with anti-biotinylated binding IgG secondary antibodies for at room temperature for 30 min. All immunohistochemical images were taken from a Nikon TE-2000 microscope, and three regions of each section were captured for semi-quantitative analysis using FIJI ImageJ (NIH). For BSP, collagen type I, and OC staining, percentage of positively stained tissue area to the whole tissue area were utilized on osteogenesis analysis. For CD 31 staining, the amount of blood vessels that formed on tissues were calculated for angiogenesis analysis47 (link),48 (link). All the other chemicals which were used in the characterizations were purchased from Vector Laboratories (CA, US).
4
Immunohistochemical Analysis of Femoral Heads
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After decalcification and paraffin embedding, femoral heads were sectioned at a thickness of 5 µm in the coronal plane. Some of these sections were stained with hematoxylin and eosin (H&E) to evaluate the trabecular structure, while the others were deparaffinized, antigen retrieved, incubated with anti-OC (Abcam), anti-Runx2 (Abcam) and anti-VEGF (Boshide, Wuhan, China) primary antibodies and then incubated with the appropriate biotinylated secondary antibodies. Sections were colored with DAB and counterstained with hematoxylin. Photomicrographs were acquired using a LEICA DM 4000. Then, the images of immunohistochemical staining were analyzed with the software Image-Pro Plus, with which the integrated option density (IOD) of target protein and the total area of trabecular bones were measured, and the mean density (IOD/area) was calculated and counted.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!