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Western blot showing reactivity of rabbit serum from different sources such as (a). rabbit anti-rohu IgM serum, (b). normal rabbit serum (from Institute's animal house), (c). normal rabbit serum (Abgenex, Bhubaneswar, India), (d). rabbit IgG against rohu lysoG (Abgenex, Bhubaneswar, India), (e). rabbit serum to MrNV protein (ICAR-CIBA, Chennai, India), and (f). normal guinea pig serum with Argulus antigens.

Western blot showing reactivity of rabbit serum from different sources such as (a). rabbit anti-rohu IgM serum, (b). normal rabbit serum (from Institute's animal house), (c). normal rabbit serum (Abgenex, Bhubaneswar, India), (d). rabbit IgG against rohu lysoG (Abgenex, Bhubaneswar, India), (e). rabbit serum to MrNV protein (ICAR-CIBA, Chennai, India), and (f). normal guinea pig serum with Argulus antigens.

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Article
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A study was undertaken to develop a western blot method for detection of immunogenic proteins of fish ectoparasite, Argulus siamensis for its further use as potential vaccine candidates. Argulus antigens were prepared by homogenization and injected to rohu (Labeo rohita) juveniles for development of immune serum. The serum was used to immunostain t...

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Context 1
... western blot was run and developed in a similar fashion without adding rohu serum samples. The results showed strong positive reaction of Argulus anti- gens with rabbit anti-rohu IgM serum sample (Figure 3(a)). Hence, the experiment was repeated with a number of rabbit serum samples from a variety of sources in order to verify whether the cross-reaction was specific to the rabbit anti-rohu IgM serum used in the test. ...
Context 2
... rabbit anti-rohu IgM serum sample (Figure 3(a)). Hence, the experiment was repeated with a number of rabbit serum samples from a variety of sources in order to verify whether the cross-reaction was specific to the rabbit anti-rohu IgM serum used in the test. The result clearly depicted the reaction of rabbit serum samples with Argulus antigens ( Figure. 3(b-e)) With an objective to remove rabbit serum from the test system, one normal guinea pig serum instead of rabbit serum was also tested and developed with goat anti-guinea pig IgG ALP conjugate and substrate. This serum sample did not show any reaction with Argulus antigens (Figure ...
Context 3
... With an objective to remove rabbit serum from the test system, one normal guinea pig serum instead of rabbit serum was also tested and developed with goat anti-guinea pig IgG ALP conjugate and substrate. This serum sample did not show any reaction with Argulus antigens (Figure 3(f)). ...

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... Over the years, ICAR-CIFA has developed a challenge facility for year-round availability of Argulus parasite and standardized challenge protocol. In another attempt, a western blot method has also been developed and used to identify immunogenic proteins of Argulus that could be targeted as potential vaccine candidates [34]. The ICAR-CIFA has interdisciplinary research collaborations, proper amenities for wet lab maintenance, skilled manpower, basic instruments, and molecular biology laboratory facility with widely experienced researchers to address the challenges faced in fish louse Argulus management. ...
... Two µl of Argulus homogenate sample containing 3 µg of protein concentration was placed onto two separate nitrocellulose membranes each. The samples were allowed to dry on the membranes, blocked with 5% skim milk (prepared in TBS) for 2 h and developed as per Das et al18 In brief, the membranes were incubated sequentially with rohu serum (Argulus immunized serum or control serum in 1:500 dilution), guinea pig anti-rohu IgM serum gel electrophoresis and Western blotting of Argulus antigens 2.4.1 | Preparation of protein samples from Argulus parasites Live Argulus parasites were collected in an Eppendorf tube and washed thoroughly with Millipore water by centrifugation. Approximately 50 numbers of parasites were homogenized in 0.5 ml of Millipore water along with 10 µl of protease inhibitor cocktail (Promega) with the help of Super FastPrep-1 homogenizer (MP Biomedicals) using lysing matrix C at a speed setting of 25 (4000 cycles per min) for 10 s. ...
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Aim An immunoproteomic approach was followed to identify immunoreactive antigens of fish ectoparasite, Argulus siamensis with rohu (Labeo rohita) immune sera for screening of potential vaccine candidates. Materials and results The whole adult Argulus antigen was run in 2D electrophoresis with IEF in 7 cm IPG strips of pH 4‐7 and SDS‐PAGE with 12% acrylamide concentration. Two parallel gels were run; one was stained with silver stain, and the other was western blotted to nitrocellulose paper (NCP) and reacted with rohu anti‐A. siamensis sera. Fourteen protein spots corresponding to the spots developed in NCP were picked from the silver‐stained gel and subjected to mass spectrometry in MALDI‐TOF/TOF. The MS/MS spectra were analyzed in MASCOT software with taxonomy ‘other metazoa’ and the proteins identified based on similarity with the proteins from heterologous species. The gene ontology analysis revealed a majority of proteins being involved in binding activity in ‘molecular function’ and belonging to metabolic processes in ‘biologic process’ categories. The possibility of these proteins as vaccine candidates against A. siamensis is discussed in the paper. Conclusion Three of the identified proteins namely, bromodomain‐containing protein, anaphase‐promoting complex subunit 5, and elongation factor‐2 could possibly serve as vaccine candidates against argulosis in carps.
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