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A Novel Monoclonal Antibody Against a Synthetic Peptide From β-Actin Can React With Its Corresponding Protein

A Novel Monoclonal Antibody Against a Synthetic Peptide From β-Actin Can React With Its Corresponding Protein

Nazila Amini, Ali-Ahmad Bayat, Omid Zarei, Reza Hadavi, Jafar Mahmoudian, Ahmad R Mahmoudi, Maryam Darzi, Hodjattallah Rabbani, Mahmood Jeddi-Tehrani

Protein Pept Lett. 2015;22(5):419-24.

PMID: 25552314

Abstract:

Actin is one of the most widely studied structural and multifunctional housekeeping proteins in eukaryotic cells with important roles in many cell functions. Antibodies against β-actin and other housekeeping gene-encoded proteins are used as internal loading controls in Western blot analyses. The aim of this study was to produce a monoclonal antibody (mAb) against a synthetic peptide derived from N-terminal region of β-actin and to study its reactivity with different organisms. A synthetic peptide, derived from β-actin, was designed and used to produce a mAb by hybridoma technology. The produced antibody (clone 4E5- A10) was purified by an affinity chromatography column followed by characterization of purified mAb using SDS-PAGE, ELISA and Western blot. Our results showed that 4E5-A10 was an IgM and had desired purity and excellent reactivity with the immunizing peptide with an affinity constant of 2.7x10(8) M(-1)>. It could detect a band of about 45 kDa, corresponding to β-actin, in Western blot. Furthermore, it could react in a more sensitive manner and with a wider range of organisms than a known commercial anti β-actin antibody. Our data suggest that 4E5-A10 can act as a sensitive probe for detection of β-actin as an internal loading control, for a wide range of organisms, in Western blot analyses.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42413877	Anti-β-Actin antibody, Mouse monoclonal			Price
IAR42413888	Monoclonal Anti-β-Actin antibody produced in mouse			Price

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