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Ab254015

Manufactured by Abcam
Sourced in United Kingdom, United States

Ab254015 is a laboratory product offered by Abcam. It is a core laboratory tool, but a detailed description cannot be provided while maintaining an unbiased and factual approach.

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2 protocols using ab254015

1

Cultured Endometrial Cell Immunohistochemistry

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The cultured endometrial cells were inoculated on sterile coverslips at 1 × 105/ml and cultured in an atmosphere of 5% CO2 at 37°C for 5 days, washed three times with PBS, soaked in methanol at −20°C for 10 min, and blocked with 5% normal goat serum (NGS) at 37°C for 30 min. The cells were incubated overnight at 4°C with a primary antibody: keratin (ab51054; 1 : 100; Abcam, Cambridge, United Kingdom) for glandular epithelial cells, vimentin (ab254015; 1 : 100; Abcam) for stromal cells, anti-LHR (sc-25828; 1 : 50; Santa Cruz Biotechnology, Santa Cruz, CA, USA), annexin IV (3175–1; 1 : 500; RabMabs, Burlingame, CA, USA), and VEGF (sc-705; 1 : 50; Santa Cruz). PBS, instead of the primary antibody, was used for negative control. The secondary antibody was HRP-linked anti-mouse IgG (7076S; 1 : 1500; Cell Signaling Technology, Inc., Danvers, MA, USA) for 30 min at room temperature. The staining was revealed with DAB for 3–5 min. The cells were counterstained with hematoxylin for 30 s. The cells were dehydrated and mounted with a mounting medium (Wuhan Boshide Biotechnology Co., Wuhan, China). Examination and photography were performed using an inverted microscope (Carl Zeiss GmbH, Oberkochen, Germany).
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2

Cultivating and Characterizing NPDFs

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Patients with NPs were recruited, and NPDFs were cultivated as previously reported [49 (link)]. The study was approved by the local ethics committee of Inje University, Busan Paik Hospital, Busan, Republic of Korea (Approval code: 80/2020). The purity of the NPDFs was checked against a panel of antibodies of fibroblast marker (cat. No. ab254015, Abcam Inc., Cambridge, MA, USA) and characteristic cell morphology data. NPDFs were used for 4th to 6th cell passages.
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