A Systematic Review: Comparison of Immunocytochemistry, ELISA, and Western Blot Methods in Alkaline phosphatase Measurement at Genistein-induced Osteoblast Cell

Burhan Ma’arif¹ , Iffatul Abada¹, Anisah Mahardiani¹, Abdul Hakim^1*, Novia Maulina¹, Neny Purwitasari², Khoirul Hidayah³ and Seow Lay Jing⁴

¹Department of Pharmacy, Faculty of Medical and Health Science, Maulana Malik Ibrahim State Islamic University, Malang, Indonesia

²Department of Pharmaceutical Science, Faculty of Pharmacy, Universitas Airlangga, Surabaya, Indonesia

³Faculty of Sharia, Maulana Malik Ibrahim State Islamic University, Malang, Indonesia

⁴Department of Pharmaceutical Chemistry, Faculty of Pharmacy and Health Sciences, Universiti Kuala Lumpur Royal College of Medicine, Perak, Malaysia

Corresponding Author E-mail: ahakim@farmasi.uin-malang.ac.id

Abstract: Osteoporosis is a bone disorder characterized by the decrease of bone mass along with bone micro-architecture damage and has a risk become a fracture. One of the causes of osteoporosis is estrogen deficiency. Genistein is a phytoestrogen compound in the isoflavone group containing a similar structure compared to 17β-estradiol, thus it can bind to estrogen receptors and produce an estrogenic effect. Genistein induction can stimulate bone formation and promote the increase of alkaline phosphate (ALP) activities in osteoblast cells which can be observed by immunocytochemistry or Enzyme-linked Immunosorbent Assay (ELISA) or Western blot method. Using the PRISMA guideline technique, choose and strategize article searches by reading the title, abstract, and then the whole text of the article. Articles with the keywords "genistein or osteoblast cells or alkaline phosphate or immunocytochemistry or immunofluorescence or ELISA or western blot" were retrieved from databases including Google Scholar, PubMed, Researchgate, and Sciencedirect. 24 relevant research articles were uncovered as a result of this systematic review. Comparison of immunocytochemistry and ELISA methods in order to analyze the activities of ALP in osteoblast induced by genistein includes selectivity, sensitivity, processing time, and cost efficiency parameters. The immunocytochemistry method has a higher level of sensitivity and a faster processing time, whereas the ELISA method has a higher level of selectivity and less cost efficiency. The western blot method has selectivity for detecting complex-level protein expression.

Keywords: Cost Efficiency; ELISA; Immunocytochemistry; Selectivity; Sensitivity, Processing Time; Western Blot

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