Paraffin embedded sections of human and murine tissues were subjected to haematoxylin and eosin (H&E) or immunofluorescence staining with antibodies directed against keratin 5 (rabbit, Abcam), CADM1 (chicken, MBL), E-cadherin (HECD1, mouse, Abcam), BrdU (rat, Abcam), Luciferase (rabbit, Abcam), keratin 14 (rabbit, Covance), and phospho-STAT3 (Cell Signalling). Detection of apoptotic cells was via TUNEL staining (Promega). Immunostained cells were fixed using 4% paraformaldehyde and incubated with antibodies to CADM1 (chicken, MBL), ITGA6 (rabbit, R&D), and HER2 (Rabbit, Cell Signalling).
Keratin 5
Manufactured by Abcam
Keratin-5 is a structural protein that is a major component of intermediate filaments in epithelial cells. It plays a crucial role in the formation and maintenance of the cytoskeleton.
Automatically generated - may contain errors
Lab products found in correlation
Keratin 8, by Abcam (2 mentions)
Apotome, by Zeiss (2 mentions)
Bromodeoxyuridine (brdu), by Abcam (1 mentions)
Tunel staining, by Promega (1 mentions)
Phospho stat3, by Cell Signaling Technology (1 mentions)
Keratin 14, by Fortrea (1 mentions)
E cadherin, by Abcam (1 mentions)
Claudin 3, by Thermo Fisher Scientific (1 mentions)
Apotome widefield microscope, by Zeiss (1 mentions)
Goat igg hrp conjugated antibody, by R&D Systems (1 mentions)
4 protocols using keratin 5
1
Multimodal Tissue Immunophenotyping
2
Thymic Microarchitecture Immunofluorescence
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Thymi were harvested and embedded in Optimal Cutting Temperature (OCT) media (Tissue-Tek). 8μm frozen thymic sections were fixed in 100% acetone, blocked in 10% goat serum, and then stained for keratin-5 (Abcam), keratin-8 (Abcam), claudin-3 (Invitrogen), or Aire (eBiosciences). Secondary antibodies were purchased from Invitrogen. Immunofluorescent staining was visualized using a Zeiss Apotome widefield microscope.
3
Analyzing Thymic and Tumor Sections
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Thymi were harvested and embedded in O.C.T. media (Tissue-Tek). 8-µm frozen thymic sections were fixed in 100% acetone, blocked, and stained for keratin-5, keratin-8 (Abcam), or Aire (eBioscience). Secondary antibodies were purchased from Invitrogen. Thymic sections were visualized using a widefield microscope (Apotome; Carl Zeiss). For tumor immunostaining, tumors were harvested and fixed in 10% formalin as previously described (Zhu et al., 2013 (link)). Fixed tumors were embedded in paraffin and sectioned for staining with anti-CD3 antibody and counterstained with DAPI. Tumor sections were visualized using a fluorescent microscope (BX60; Olympus) and analyzed using ImageJ software (National Institutes of Health).
4
Immunohistochemical Analysis of Organoid Tissues
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Organoids and tissue were fixed in 4% paraformaldehyde for 1–6 h, dehydrated, and paraffin-embedded according to standard histology procedures. Sections were stained with Hematoxylin & Eosin (HE) or the following antibodies: Keratin 5 (Abcam, ab52635), Ki67 (Dako, M7240), TP53 (Santa Cruz, sc-126), TP63 (Abcam, ab735) and Uroplakin III (SFI-1, Abcam, ab78196) according to the manufacturer’s protocols. For Keratin 20 (DakoCytomation, M7019), EDTA antigen retrieval was used in combination with a Ultravision Protein Block (Fisher Scientific, TA-125-PBQ) for 30 min at room temperature. The following secondary antibodies conjugated with HRP were used: bright vision poly-hrp-anti-rabbit igg (VWR international, VWRKDPVR110HRP) and bright vision poly-hrp-anti-mouse igg (VWR international, VWRKDPVM110HRP) for 1 h. at room temperature, and Goat IgG HRP-conjugated Antibody (R&D Systems Europe, HAF017) for 1 h. at 37 °C. Staining was performed using 3,3’-diaminobenzidine (DAB) for 10 min exactly at room temperature. Images were acquired by high-resolution scanning of the slides and analysed using the NDP.view (v2.7.39) software (Hamamatsu Photonics K.K.). These sections were evaluated in a blinded test by dedicated uropathologists. Regrettably, due to a clerical error, the original tissue of UBTOR5 was not available for immunohistochemical analysis and thus excluded for the comparison.
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