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Brucella polyclonal rabbit antibody

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The Brucella polyclonal rabbit antibody is a laboratory reagent used for the detection and identification of Brucella species in various applications. It is produced by immunizing rabbits with Brucella antigens, resulting in a polyclonal antibody preparation.

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Lab products found in correlation

Normal goat serum (ngs), by Vector Laboratories (3 mentions) Bloxall blocking solution, by Vector Laboratories (3 mentions) Betazoid dab chromagen kit, by Biocare Medical (3 mentions) Ems solution a, by Electron Microscopy Sciences (2 mentions) Vectastain abc, by Biocare Medical (2 mentions) Vectastain elite abc hrp kit, by Vector Laboratories (1 mentions)

4 protocols using brucella polyclonal rabbit antibody

1

Immunohistochemical Detection of Brucella in Tissues

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Unstained slides from liver, spleen, uterus, and mesenteric lymph nodes were adhered to positively charged glass slides for immunohistochemistry. Slides were deparaffinized and rehydrated through a series of xylene and ethanol steps before antigen retrieval was performed using 1:10 EMS Solution A (Electron Microscopy Services) in a 2100 Antigen Retriever (Aptum Biologics Ltd.), according to the manufacturer’s instruction. Endogenous peroxidase activity was blocked by 10 m incubation with Bloxall Blocking Solution (Vector Laboratories) followed by 20 m blocking with normal goat serum (Vector). After each step slides were washed with PBS plus 0.5% tween for 5 minutes. Primary incubation was overnight at 4°C with Brucella polyclonal rabbit antibody (Bioss) at 1:400. Negative control tissues were incubated with rabbit nonimmune serum diluted in PBS. A Vectastain ABC and Betazoid DAB chromagen kits (Biocare Medical) were used following primary incubation according to the manufacturer’s instructions. The slides were counterstained with Meyer’s hematoxylin III.
Stranahan L.W., Khalaf O.H., Garcia-Gonzalez D.G, & Arenas-Gamboa A.M. (2019). Characterization of Brucella canis infection in mice. PLoS ONE, 14(6), e0218809.
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2

Histopathological Analysis of Vaccination Effects

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To assess of histologic changes associated with vaccination, multiple tissues were collected at necropsy from moms, their offspring, and bucks (Table S1). Tissues were fixed in 10% neutral buffered formalin and were then routinely processed and embedded in paraffin, sectioned at 5 μm, and stained with hematoxylin and eosin. Histologic changes between groups were evaluated in a blinded fashion by a boarded veterinary anatomic pathologist. Unstained sections from placenta were adhered to positively charged glass slides for immunohistochemistry. Following deparaffinization and rehydration through a series of xylene and ethanol steps, antigen retrieval and blocking was performed as previously described [11 (link)]. Primary incubation was done overnight at 4° C with Brucella polyclonal rabbit antibody (Bioss, Boston, MA) at 1:2,000 ratio. Negative control tissues were incubated with rabbit nonimmune serum diluted in PBS. Vectastain ABC and Betazoid DAB chromogen kits (Biocare Medical, CA) were used following primary incubation according to the manufacturer’s instructions. The slides were counterstained with Meyer’s hematoxylin III.
Castaño-Zubieta M.R., Rossetti C.A., García-González D.G., Maurizio E., Hensel M.E., Rice-Ficht A.C., Ficht T.A, & Arenas-Gamboa Á.M. (2020). Evaluation of the safety profile of the vaccine candidate Brucella melitensis 16MΔvjbR strain in goats. Vaccine, 39(3), 617-625.
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3

Immunohistochemical Detection of Brucella in Tissues

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Unstained slides from spleen, uterus, liver, and lung were adhered to positively charged glass slides for immunohistochemistry. Slides were deparaffinized and rehydrated through a series of xylene and ethanol steps before antigen retrieval was performed using 1:10 EMS Solution A (Electron Microscopy Services) in a 2100 Antigen Retriever (Aptum Biologics Ltd.), according to manufacturer protocol. Endogenous peroxidases were blocked by 10 m incubation with Bloxall Blocking Solution (Vector Laboratories) followed by 20 m blocking with normal goat serum (Vector). After each step slides were washed with PBS plus 0.5% tween for 5 minutes. Primary incubation was overnight at 4o C with Brucella polyclonal rabbit antibody (Bioss) at 1:600. Negative control tissues were incubated with rabbit nonimmune serum diluted in PBS. A Vectastain ABC and Betazoid DAB chromagen kits (Biocare Medical) were used following primary incubation according to the manufacturer’s instructions. The slides were counterstained with Meyer’s hematoxylin III.
Hensel M.E., Garcia-Gonzalez D.G., Chaki S.P., Samuel J, & Arenas-Gamboa A.M. (2019). Characterization of an intratracheal aerosol challenge model of Brucella melitensis in guinea pigs. PLoS ONE, 14(3), e0212457.
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4

Immunohistochemical Detection of Brucella

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Five micrometer tissue sections of testicle/epididymis (mouse and guinea pig) and prostate (guinea pig only) were adhered to positively charged glass slides for immunohistochemistry. Slides were routinely processed, and antigen retrieval was performed as previously described using a 2,100 Antigen Retriever (Aptum Biologics Ltd. Southampton, UK; Hensel et al., 2019 (link)). Slides were blocked as previously described with Bloxall Blocking Solution (Vector Laboratories, Burlingame, CA) and normal goat serum (Vector Laboratories; Hensel et al., 2019 (link)). Primary incubation was performed overnight at 4o C with a Brucella polyclonal rabbit antibody (Bioss Antibodies, Boston, MA) at dilution of 1:500. A Vectastain Elite® ABC HRP Kit (Vector Laboratories) with an avidin/biotinylated anti-rabbit secondary antibody was used according to the manufacturer’s instructions. Antigen was visualized with a Betazoid DAB chromagen kit (Biocare Medical, Pachecho, CA). The slides were counterstained with Gills’s hematoxylin III and cover slipped.
Hensel M.E., Stranahan L.W., Edwards J.F, & Arenas-Gamboa A.M. (2022). Intratracheal inoculation results in Brucella-associated reproductive disease in male mouse and guinea pig models of infection. Frontiers in Microbiology, 13, 1029199.
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