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Table 3 Antibodies used in this study

From: Histopathological findings, phenotyping of inflammatory cells, and expression of markers of nitritative injury in joint tissue samples from calves after vaccination and intraarticular challenge with Mycoplasma bovis strain 1067

Antibody designation

Type or isotype

Specificity

Dilution

Antigen retrieval

References/source

D490

Rabbit hyperimmune serum

M. bovis

1:1800

0.25% trypsina (37°C, 60 min)

[26]

mAbb pool

Mouse IgG and IgM

M. bovis

1:2000

0.25% trypsin (37°C, 60 min)

Chemicon

1A1

Mouse IgG1

M. bovis VspA, C, and other Vspsc

1:200

0.25% trypsin (37°C, 60 min)

[27]

1E5

Mouse IgM

M. bovis VspA, B, C

undiluted

0.25% trypsin (37°C, 60 min)

[26],[28],[29]

Anti-CD3

Rabbit IgG

CD3+ T lymphocytes

1:300

Demasking solution Gd (95°C,

DakoCytomation

BAQ44A

Mouse IgM

B lymphocytes

1:100

20 min) 0.01 M citric buffer (microwave 95°C, 10 min)

VMRD; [23]

Anti-MRP8 (S100A8)

Mouse IgG

Human MRP8e

1:600

Pronase Ef (37°C, 20 min) [26],

BMA Biomedicals; [30]

Anti-MRP14 (S100A9)

Mouse IgG

Human MRP14g

1:8000

0.01 M citrate buffer (microwave 95°C, 10 min)

BMA Biomedicals; [30]

TAL.1B5

Mouse IgG1

α-chain of human leukocyte antigen (HLA-DR)

1:4000

0.01 M citrate buffer (microwave 95°C, 15 min)

DakoCytomation

Anti-iNOS/NOS II

Rabbit IgG

iNOS

1:50

Saponinef (37°C, 20 min)

[31],[32]; Biomol GmbH

Anti-NT

Rabbit IgG

NT

1:100

Pronase E (37°C, 20 min)

Millipore GmbH

  1. aFluka, Buchs, Switzerland. bmonoclonal antibody. cvariable surface proteins belonging to the Vsp protein family of M. bovis.
  2. dBIOLOGO, Kronshagen, Germany. eCalgranulin A fVWR TM International GmbH, Darmstadt, Germany. gCalgranulin B.