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Ab218527

Manufactured by Abcam
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Sourced in United States

Ab218527 is a lab equipment product from Abcam. The core function of this product is to [description not available].

Automatically generated - may contain errors

Lab products found in correlation

Ab235958, by Abcam (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Ab8245, by Abcam (1 mentions) Ab81283, by Abcam (1 mentions) Ab54230, by Abcam (1 mentions) Ab201015, by Abcam (1 mentions) Ab13939, by Abcam (1 mentions) Ab40866, by Abcam (1 mentions) Ab6721, by Abcam (1 mentions) Ab185622, by Abcam (1 mentions) β actin, by Santa Cruz Biotechnology (1 mentions) Ab6728, by Abcam (1 mentions) Sc 5294, by Santa Cruz Biotechnology (1 mentions) Sc 25778, by Santa Cruz Biotechnology (1 mentions) Ab128040, by Abcam (1 mentions) Ab191387, by Abcam (1 mentions)

4 protocols using ab218527

1

Protein Extraction and Western Blot Analysis

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Protein was obtained from cells with RIPA lysis buffer (Beyotime, Shanghai, China) supplied with protease inhibitors. Protein was separated through SDS-PAGE and moved to PVDF membranes (Millipore, Darmstadt, Germany). After being sealed with 5% skimmed milk, the membranes were cultivated with primary antibodies for E2F1 (ab218527, 1/1000), SYTL2 (ab231133, 1/1000), Akt (ab235958, 1/1000), p-Akt (ab81283, 1/5000), ERK (ab54230, 1/1000), p-ERK (ab201015, 1/1000) and GAPDH (ab8245, 1/1000) from Abcam (Cambridge, USA). Secondary antibodies were added for cultivating for 1 h. The amount of protein was examined via chemiluminescence detection system.
Zhou X., Li J., Teng J., Liu Y., Zhang D., Liu L, & Zhang W. (2020). Long noncoding RNA BSN-AS2 induced by E2F1 promotes spinal osteosarcoma progression by targeting miR-654-3p/SYTL2 axis. Cancer Cell International, 20, 133.
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2

Quantitative Immunoblotting of Cell Cycle Proteins

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Cells were seeded into 6 cm plates (4 × 105 cells/plate) for 48 h and the lysates were subjected to SDS-PAGE electrophoresis, transferred onto a nitrocellulose membrane, and blocked with 5% skim milk at room temperature. The membranes were incubated with primary antibodies overnight, washed with phosphate buffered saline (PBS), and incubated with anti-mouse (Abcam, ab6728) or rabbit IgG-HRP (Abcam, ab6721). The primary antibodies were as follows: E2F1 (Abcam, ab218527), AURKB (Abcam, ab24), DSN1 (Proteintech, 17,742–1-AP), CCNB2 (Abcam, ab185622), CHEK1 (Abcam, ab40866), CENPA (Abcam, ab13939), DP1 (Abcam, ab124678), β-actin (Santa Cruz Biotechnology, sc69879), and GAPDH (Proteintech, 10,494–1-AP).
Wang H., Yu S., Peng H., Shu Y., Zhang W., Zhu Q., Wu Y., Xu Y., Yan J, & Xiang H. (2020). Long noncoding RNA Linc00337 functions as an E2F1 co-activator and promotes cell proliferation in pancreatic ductal adenocarcinoma. Journal of Experimental & Clinical Cancer Research : CR, 39, 216.
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3

Cellular Protein Extraction and Immunoblotting

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Cellular protein was extracted as described previously [23 (link)]. Antibodies against E2F1 (ab218527, Abcam, Cambridge, MA, USA), TXNIP (ab188865), ASK1 (sc-5294, Santa Cruz, CA, USA) and GAPDH (sc-25778) were used. GAPDH served as the internal control. ImageJ v1.50e was used to quantify the band intensity.
Gao S., Bu X., Gao Y., Bao Z., Shi W., Luan L., Chen H., Zhang B., Tian Q., Guan W, & Yang L. (2022). The miR-532-E2F1 feedback loop contributes to gastric cancer progression. Cell Death & Disease, 13(4), 376.
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4

Immunohistochemical Analysis of Cartilage Markers

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Clinical samples were collected and processed to isolate cartilage tissues, and the tissue sections were subjected to immunohistochemical detection of KDM2A-, E2F1-, and PTTG1-positive expression rates. Antigens were retrieved from tissue sections by boiling in sodium citrate buffer. Immunohistochemistry was carried out by incubating sections with primary antibodies rabbit anti-KDM2A (ab191387; Abcam, Cambridge, UK), E2F1 (ab218527; Abcam, Cambridge, UK), and PTTG1 (ab128040; Abcam, Cambridge, UK). Normal rabbit serum was employed as NC instead of primary antibody. Next, the sections were incubated with HRP-secondary antibody and then exposed to 3,3′-diaminobenzidine (DAB) reagent prior to microscopic observation. KDM2A, E2F1, and PTTG1 expression was positive with brownish-yellow particles in chondrocytes. Specimens were scored according to the intensity of the dye color and the number of positive cells.38 (link)
Wang K., Li F., Yuan Y., Shan L., Cui Y., Qu J, & Lian F. (2020). Synovial Mesenchymal Stem Cell-Derived EV-Packaged miR-31 Downregulates Histone Demethylase KDM2A to Prevent Knee Osteoarthritis. Molecular Therapy. Nucleic Acids, 22, 1078-1091.
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