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Ab97021

Manufactured by Abcam
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Ab97021 is a laboratory equipment product designed for general research purposes. It is a device that serves a core function without any interpretations or extrapolations on its intended use.

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Lab products found in correlation

Ab90395, by Abcam (1 mentions) C7974, by Merck Group (1 mentions) Digest all, by Thermo Fisher Scientific (1 mentions) Ii ii6b3, by Developmental Studies Hybridoma Bank (1 mentions) Ab26278, by Abcam (1 mentions) Ab64238, by Abcam (1 mentions) Rodent block m rbm961g, by Biocare Medical (1 mentions)

3 protocols using ab97021

1

Histological Analysis of iPSC Cartilage

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After three days of culture, iPSC cartilage and native cartilage samples were
fixed in 4% paraformaldehyde and embedded in paraffin. Sections of 8 μm
were stained with safranin-O/fast green/hematoxylin or picrosirius red.
Immunohistochemistry for collagen type II was performed as previously described (27 (link)) using a primary antibody from the Developmental
Studies Hybridoma Bank (II-II6B3, 1:1), a secondary antibody form Abcam (ab97021, 1:500),
and the AEC Red Single chromogen (Invitrogen). Two samples per group were used for
histological analysis.
Willard V.P., Diekman B.O., Sanchez-Adams J., Christoforou N., Leong K.W, & Guilak F. (2014). Use of cartilage derived from murine induced pluripotent stem cells for osteoarthritis drug screening. Arthritis & rheumatology (Hoboken, N.J.), 66(11), 3062-3072.
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2

Histological Analysis of Engineered Cartilage

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Constructs were fixed in 4% paraformaldehyde with 100 mM sodium cacodylate overnight. Subsequently, constructs were dehydrated through a series of ethanol washes, cleared in xylenes, embedded in paraffin, and sectioned at 8 μm per section. Sections from each sample were stained for GAGs with 0.1% aqueous Safranin-O, collagen with 0.02% aqueous fast green, and stained for cell nuclei with hematoxylin. Immunohistochemistry was performed with monoclonal antibodies to type I collagen (ab90395; Abcam, Cambridge, MA), type II collagen (II-II6B3; Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), or type X collagen (C7974; Sigma-Aldrich). For all IHC, slides with sections of each sample were cleared, rehydrated, and digested with pepsin for epitope retrieval (Digest-All, Invitrogen). After probing with appropriate primary antibodies, all samples were probed with the same biotinylated anti-mouse secondary antibody (ab97021; Abcam), treated with HRP conjugate (Invitrogen), and finally incubated with chromagenaminoethylcarbazole (AEC) single solution (Invitrogen). Human osteochondral plugs were used as positive controls for each antibody and for the general histological stain. Negative controls without primary antibody were also prepared for each slide.
Glass K.A., Link J.M., Brunger J.M., Moutos F.T., Gersbach C.A, & Guilak F. (2014). Tissue-engineered cartilage with inducible and tunable immunomodulatory properties. Biomaterials, 35(22), 5921-5931.
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3

Immunohistochemical Analysis of GLP-1 Expression

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Segments of distal ileum (1 cm of intestine withdrawn above the junction with the caecum) were harvested from all rats and fixed in 10% neutral-buffered formalin for 24 h, dehydrated and embedded in paraffin. The paraffin-embedded tissue sections were cut (3 μm thickness) and mounted on adhesive microscope slides. The sections were deparaffinized in xylene and rehydrated with decreasing concentrations of ethanol (100%, 95% and 70%) and distilled water. Antigen retrieval was performed in 0.01 M sodium citrate buffer with 0.05% Tween 20 (pH 6.0), using a water bath at 98 °C for 10 min and blocked in 3% H2O2 in PBS. After that, sections were incubated with a blocking solution (Rodent Block M, RBM961G, Biocare Medical, UK) for 30 minutes and then incubated overnight at 4 °C with anti-GLP-1 primary antibody: mouse monoclonal, ab26278; Abcam, Amsterdam, The Netherlands; 1:1500. Subsequently, sections were incubated with secondary antibody at RT for 30 min: goat anti-mouse, ab97021; Abcam, Amsterdam, The Netherlands; 1:200. Then, 30-minute ABC complex incubation was carried out (Vector A and B, Vectastain, Vector Labs, Peterborough, UK). Finally, the sections were treated with diaminobenzidine tetrahydrochloride (DAB) substrate kit (ab64238, Abcam, Amsterdam, The Netherlands). All sections were stained with H&E.
Pena M.J., Costa R., Rodrigues I., Martins S., Guimarães J.T., Faria A., Calhau C., Rocha J.C, & Borges N. (2021). Unveiling the Metabolic Effects of Glycomacropeptide. International Journal of Molecular Sciences, 22(18), 9731.
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