Biomedical Chemistry: Research and Methods 2024, 7(4), e00248

Interaction of Mouse and Sheep Polyclonal Antibodies with the Main Forms of Human and Rat Renalase

V.I. Fedchenko*, A.A. Kaloshin, S.A. Kaloshina, A.E. Medvedev

Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia; *e-mail: valfed38@yandex.ru

Keywords: polyclonal antibodies; human renalase-1 (RNLS1-human); rat renalase-2 (RNLS2-rat)

DOI:10.18097/BMCRM00248

The whole version of this paper is available in Russian.

The interaction of sheep and mouse polyclonal antirenalase antibodies obtained by immunization with full-length human (RNLS1-human) and rat (RNLS2-rat) renalases, respectively, has been studied. The target recombinant proteins, RNLS1-human and RNLS2-rat, were expressed in E. coli cells and isolated by Ni-agarose chromatography. Sheep polyclonal antibodies against RNLS1-human interacted more effectively with both RNLS1-human than with RNLS2-rat. Mouse polyclonal antibodies against RNLS2-rat effectively interacted mainly with RNLS2-rat, but not with RNLS1-human. The data obtained indicate the preferential selectivity of the antibody interaction with the proteins against which they were obtained. This should be taken into consideration in the case of selection of commercially available antibody preparations for quantitative immunodetection of target proteins in biological objects.
Figure 1. A - electrophoresis in 12% SDS-PAAG of the recombinant proteins RNLS1-human and RNLS2-rat after transfer to a nitrocellulose membrane and Ponceau S staining. 1) RNLS1-human.
2) RNLS2-rat. M) - protein molecular mass markers of 75 kDa, 50 kDa, 37 kDa, 25 kDa, and 20 kDa.
B - Western blot analysis of the interaction of polyclonal antibodies against RNLS1-human with RNLS1-human and RNLS2-rat.
C - Western blot analysis of the interaction of polyclonal antibodies against RNLS2-rat with RNLS1-human and RNLS2-rat.

Figure 2. Blast analysis of the amino acid sequences of RNLS1-human and RNLS2-rat. RNLS1-human – amino acid sequence of human renalase-1. RNLS2-rat – amino acid sequence of rat renalase-2. Identical amino acid residues of RNLS1-human and RNLS2-rat shown between RNLS1-human and RNLS2-rat are highlighted in italics.

FUNDING

The work was performed within the framework of the Program for Basic Research in the Russian Federation for a long-term period (2021-2030) (№ 122030100170-5).

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