Immunohistochemical assessment was performed according to the procedures described previously [29 (link)]. Target proteins in the cartilage were detected using the following primary antibodies: TGF-β1(1:200)(ab92486, Abcam, Inc., USA), ADAMTS-5 (1:500)(BA3020, Boster Co., Ltd., China), Col-II (1:500) (P02458, Boster), MMP-13 (1:500)(P23097, Boster), collagen type X (Col-X,1:200)(bs-0554R-FITC, Bioss Co., Ltd., China), and AGG (1:500)(GTX54920, Gene Tex, Inc., USA). In addition, collagen type I (Col-I,1: 200) (BA0325, Boster) was assayed to evaluate the level of bone matrix in the subchondral bone. The assessment of the staining was based on the average sum of the integrated optical density (IOD/mm2), which is the ratio of the total IOD and the area of the region of interest (ROI) (mm2) [29 (link)]. The cartilage of the load-bearing zone was defined as ROI [30 (link)]. Measurements were performed using Image-Pro Plus version 6.0.0.260 (Media Cybernetics, Inc., USA).
Gtx54920
Manufactured by GeneTex
Sourced in United States
GTX54920 is a compact and versatile benchtop centrifuge designed for a wide range of laboratory applications. It features a high-capacity rotor capable of handling multiple sample types simultaneously. The centrifuge operates at a maximum speed of 6,000 RPM and can generate up to 3,000 x g of centrifugal force. The unit is equipped with a user-friendly control panel for easy operation and includes safety features to ensure a secure working environment.
Automatically generated - may contain errors
Lab products found in correlation
Eclipse ti2 e inverted microscope, by Nikon (2 mentions)
Ab34712, by GeneTex (2 mentions)
Plan fluor 10x 0.30 dic l objective, by Nikon (2 mentions)
Gtx37732, by GeneTex (2 mentions)
Ab34710, by Abcam (2 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (2 mentions)
Ab92486, by Abcam (1 mentions)
Image pro plus, by Media Cybernetics (1 mentions)
Ba0325, by Boster Bio (1 mentions)
Ba3020, by Boster Bio (1 mentions)
3 protocols using gtx54920
1
Immunohistochemical Assessment of Cartilage Proteins
2
Fluorescent IHC of Collagen and Aggrecan
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Fluorescent immunohistochemistry (IHC) was performed to assess types I, II, and X collagen, and aggrecan localization. Engineered (N = 4) and native bovine ACL ligament-to-bone (N = 3) samples were fixed, demineralized as previously described in section 2.3.3 Polarized Picrosirius Red Imaging, embedded in paraffin blocks and sectioned. Sections were treated with Proteinase K to retrieve antigens, blocked with 5% goat serum and incubated overnight with primary polyclonal rabbit antibodies in 5% goat serum at 1:150 dilution for types I (Abcam AB34710), II (Abcam AB34712), and X collagen (GeneTex GTX37732), and aggrecan (GeneTex GTX54920). Negative controls were incubated overnight in 5% goat serum. All sections were then incubated with goat anti-rabbit IgG secondary antibody labeled with Alexa Fluor 488 (Invitrogen A11008) at 1:200 dilution for 2 hours and cross labeled with DAPI at 1:1000 dilution. Stained sections were imaged using a Nikon Eclipse Ti2-E inverted microscope and Nikon Plan Fluor 10x/0.30 DIC L objective to observe protein localization.
3
Fluorescent IHC of Collagen and Aggrecan
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fluorescent immunohistochemistry (IHC) was performed to assess types I, II, and X collagen, and aggrecan localization. Engineered (N = 4) and native bovine ACL ligament-to-bone (N = 3) samples were fixed, demineralized as previously described in section 2.3.3 Polarized Picrosirius Red Imaging, embedded in paraffin blocks and sectioned. Sections were treated with Proteinase K to retrieve antigens, blocked with 5% goat serum and incubated overnight with primary polyclonal rabbit antibodies in 5% goat serum at 1:150 dilution for types I (Abcam AB34710), II (Abcam AB34712), and X collagen (GeneTex GTX37732), and aggrecan (GeneTex GTX54920). Negative controls were incubated overnight in 5% goat serum. All sections were then incubated with goat anti-rabbit IgG secondary antibody labeled with Alexa Fluor 488 (Invitrogen A11008) at 1:200 dilution for 2 hours and cross labeled with DAPI at 1:1000 dilution. Stained sections were imaged using a Nikon Eclipse Ti2-E inverted microscope and Nikon Plan Fluor 10x/0.30 DIC L objective to observe protein localization.
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