Epr1473

Manufactured by Abcam

Sourced in United Kingdom

EPR1473 is a recombinant antibody that recognizes the protein EPR1473. It is produced in HEK293 cells and is suitable for use in various applications, including immunohistochemistry, Western blotting, and ELISA.

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Lab products found in correlation

Zen blue edition program, by Zeiss (1 mentions)

Close spelling variants of this product

EPR1473) to CDKN2A/p16INK4a tag (2 citations)

4 protocols using epr1473

Profiling IDO1 Expression in Tumor Samples

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IDO1 expression in 171 PTs and 40 corresponding LNMs was investigated by IHC. TMAs were prepared following published protocols (Freier et al. 2003 (link); Moratin et al. 2019 (link)).
Slides were stained anti-IDO1 monoclonal antibody (clone D5J4E, Cell Signaling Technology, Cambridge, United Kingdom) at a dilution of 1:400 following the manufacturer’s instructions and were scanned with an Axio Scan.Z1 (Zeiss, Oberkochen, Germany) for further investigation. The TMAs were scored using ZEN (blue edition) program (Zeiss) by three independent observers.
TMAs were assessed by determining the density of IDO1⁺ immune cells and distinguishing the distribution of IDO1⁺ immune cells between peritumoural and intratumoural regions.
We used a dataset previously published by Moratin et al. to evaluate the correlation between IDO1 and PD-L1/2 (Moratin et al. 2019 (link)). In situ hybridization for human papillomavirus (HPV)-DNA was performed following published protocols (Kühn et al. 2021 (link); Linxweiler et al. 2015 (link)). Immunohistochemical staining targeting p16 was performed following the manufacturer’s instructions (clone EPR1473, Abcam, Cambridge, United Kingdom).

Struckmeier A.K., Radermacher A., Fehrenz M., Bellin T., Alansary D., Wartenberg P., Boehm U., Wagner M., Scheller A., Hess J., Moratin J., Freudlsperger C., Hoffmann J., Thurner L., Roemer K., Freier K, & Horn D. (2022). IDO1 is highly expressed in macrophages of patients in advanced tumour stages of oral squamous cell carcinoma. Journal of Cancer Research and Clinical Oncology, 149(7), 3623-3635.

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Immunohistochemical Evaluation of p16 Expression

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Formalin-fixed, paraffin-embedded tumor specimens were evaluated for p16 overexpression with a rabbit monoclonal antibody that recognizes p16 (Anti-CDKN2A/p16INK4a antibody [EPR1473]: Abcam Plc, Science Park, Cambridge, England). In this study, positive p16-protein expression (designated p16 (+)) determined via immunohistochemistry (IHC) was defined as strong and diffuse nuclear, cytoplasmic staining or both in at least 70% of tumor cells. Any other pattern of p16 expression was classified as p16 (-).

Mafune A., Hama T., Suda T., Suzuki Y., Ikegami M., Sakanashi C., Imai S., Nakashima A., Yokoo T., Wada K., Kojima H, & Urashima M. (2015). Homozygous deletions of UGT2B17 modifies effects of smoking on TP53-mutations and relapse of head and neck carcinoma. BMC Cancer, 15, 205.

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Evaluating p16 Expression by IHC

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Expression of p16 was evaluated by immunohistochemistry using rabbit anti‐p16 mAb (EPR1473; Abcam) following the manufacturer’s instructions. Positive p16 expression was defined as strong diffuse nuclear and cytoplasmic staining in 70% or more of tumor cells, according to previously described criteria.²⁴ (link)

Hashida Y., Higuchi T., Matsumoto S., Iguchi M., Murakami I., Hyodo M, & Daibata M. (2021). Prognostic significance of human papillomavirus 16 viral load level in patients with oropharyngeal cancer. Cancer Science, 112(10), 4404-4417.

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Immunohistochemical Analysis of p16INK4A and p14ARF in Tissue Samples

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FFPE slices were boiled in 10 nM sodium citrate buffer (pH 6.0) or Tris–EDTA buffer (pH 9.0) for 20 min under microwave irradiation for antigen retrieval. Immunohistochemical staining was performed using the polymeric method with the ImmPRESS universal PLUS polymer kit (Vector Laboratories, UK). After blocking with 2.5% normal horse serum for 20 min at 20 °C, the sliced tissues were incubated at 4 °C overnight with an anti-human p16INK4A monoclonal antibody (1:300; EPR1473, Abcam, UK) and anti-human p14ARF polyclonal antibody (1:200; E3X6D, Cell Signaling Technology, USA). The tissue was reacted with the ImmPRESS universal polymer reagent, containing the secondary antibody, for 30 min and then treated with diaminobenzidine for p16INK4A and p14ARF staining. More than 10% of the p16INK4A expression in the cytoplasm and p14ARF expression in the nucleus was considered positive in the neoplastic and adjacent non-neoplastic epithelium.

Onozato Y., Sasaki Y., Abe Y., Sato H., Yagi M., Mizumoto N., Kon T., Sakai T., Ito M., Umehara M., Koseki A, & Ueno Y. (2021). Novel genomic alteration in superficial esophageal squamous cell neoplasms in non-smoker non-drinker females. Scientific Reports, 11, 20150.

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