The 5 µm thick sections were prepared from the formalin-fixed and paraffin embedded tissues (3 NHP and 23 CD). IHC staining was performed as previously reported (12 (link)). Specific primary antibody against SFRP2 (dilution 1:200; cat. no. ab86379; Abcam) was used. All the IHC slides were evaluated by two pathologists (Department of Pathology, Rui-Jin Hospital, Shanghai, China). The staining intensity was graded as follows: No staining, 0; weakly positive, 1; moderately positive, 2; and strongly positive, 3. The staining percentage was graded as follows: 0–25% staining, 1; 26–50% staining, 2; 51–75% staining, 3; and 76–100% staining, 4. The immunoreactive score was calculated as intensity of the staining multiplied by the percentage of positive cells, which was then categorized as low (0–6) and high (7 (link)–12 (link)) expression.
Ab86379
Manufactured by Abcam
Sourced in United States
Ab86379 is a primary antibody produced in rabbit. It is designed for use in immunohistochemistry applications.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 710 confocal microscope, by Zeiss (3 mentions)
Orb373345, by Biorbyt (1 mentions)
Supersignaltm west femto maximum sensitivity substrate, by Thermo Fisher Scientific (1 mentions)
Ne per nuclear and cytoplasmic extraction kit, by Thermo Fisher Scientific (1 mentions)
Ab9485, by Abcam (1 mentions)
Ab25235, by Abcam (1 mentions)
Ab64693, by Abcam (1 mentions)
3 protocols using ab86379
1
Immunohistochemical Evaluation of SFRP2 Expression
2
Quantifying Nuclear and Cytoplasmic β-catenin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For determining β-catenin levels in the nucleus and cytoplasm, nuclear, and cytoplasmic protein fractions of DP cells were extracted using a NE-PER Nuclear and Cytoplasmic Extraction Kit (78835; Pierce Biotechnology, United States). Total protein from DP cells, following H19 transient transfection, and dorsal skin tissues from nude mice were also extracted. The BCA method was applied for quantifying protein concentration. RIPA buffer was used to lyse cells and tissues and SDS-PAGE was conducted to separate the cellular or tissue proteins. Antibodies and dilutions used were as follows: GAPDH (Abcam, ab9485, United States, 1:5000), TBP (CST 8515S, United States, 1:1000), β-catenin (Abcam, ab32572, United States, 1:1000), DKK1 (Abcam, ab109416, United States, 1:1000), Kremen2 (Abcam, ab156007, United States, 1:1000), sFRP2 (Abcam, ab86379, United States, 1:1000), Wnt3a (1:1000, 09-162, Millipore), and LRP6 (1:1000, orb373345, Biorbyt). Bands were visualized with SuperSignalTM West Femto Maximum Sensitivity Substrate (34095, Thermo Fisher Scientific) and a Universal Hood II gel imaging and image lab 3.0 analysis system (Bio-Rad, United States).
3
Visualizing Synovial Nerve Fibers and Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Synovial tissues and DRGs were dissected and immersed into 4% paraformaldehyde in PBS for 4 h. Then explants were embedded in O.C.T. compound (Tissue-Tek, 4583) frozen next to a cryoprotection in a solution of 20% sucrose overnight in room temperature. For immunofluorescent staining, the ankle joint specimens were sagittally sectioned to obtain free-floating 50-μm-thick slices for a better vision of nerve fibers in synovial tissue of ankle joint, while DRG explants were sectioned to regular 3.5-μm-thick slices and both stained for β-III-Tubulin (Cell Signaling Technology, D71G9, 1:400), sFRP2 (Abcam, ab86379, 1:100) in Eppendorf tube. For staining of M1/M2 macrophages in synovial tissue, 3.5-μm-thick slices were obtained for CD16/32 (Abcam, ab25235, 1:20), CD206 (Abcam, ab64693, 1:100) and hematoxylin and eosin staining.
Zeiss LSM710 confocal microscope was used for confocal imaging of samples at an original magnification ×40 and Zen software.
Zeiss LSM710 confocal microscope was used for confocal imaging of samples at an original magnification ×40 and Zen software.
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!