- Description of Target:
- MOUSE ANTI BOVINE INTERLEUKIN-12:Biotin
- Gene Symbol:
- NCBI Gene Id:
- Alias Symbols:
- Tissue Tool:
- Find tissues and cell lines supported to express IL12B.
- Protein Accession #:
- Swissprot Id:
- Protein Name:
- Interleukin-12 subunit beta
- Protein Size (# AA):
- ELISA, FC
- The immunogen for anti-IL12B antibody: recombinant bovine IL-12
- Product Format:
- Purified IgG conjugated to Biotin - liquid
- Datasheets / Downloads:
- Printable datasheet for
anti-IL12B antibody- OASA00348
- Applications Info:
- ELISA : This biotin conjugate may be used as detection reagent in a sandwich ELISA assay using OASA00349 as capture reagent.
Flow Cytometry: Use 10ul of the suggested working dilution to label 106 cells of 100ul.
- Predicted Homology Based on Immunogen Sequence:
- Species Reactivity:
- Additional Information:
- Fusion Partners: Spleen cells from immunised BALB/c mice were fused with cells of the mouse SP2/0 myeloma cell line.
- Preparation: Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
Preservative Stabilisers: 0.09% - Sodium Azide
- Approx Protein Conc: IgG concentration 1.0 mg/ml
Buffer Solutions: Phosphate buffered saline pH7.4
- Reconstitution and Storage:
- Store at +4oC or at -20oC if preferred.
This product should be stored undiluted.
Storage in frost-free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
- Target Reference:
- 1. Hope, J. C. et al. (2002) Development of detection methods for ruminant interleukin (IL) - 12. J. Immunol. Methods. 266: 117 - 126.
2. Wenz, J.R. et al. (2010) Factors associated with concentrations of select cytokine and acute phase proteins in dairy cows with naturally occurring clinical mastitis. J Dairy Sci. 93: 2458-70.
3. Rinaldi, M. et al (2010) A sentinel function for teat tissues in dairy cows: dominant innate immune response elements define early response to E. coli mastitis. Funct Integr Genomics. 10: 21-38.
Product Protocol: IL12B antibody used to evaluate mediation of host immune responses after immunization of neonatal calves with a heat-killed mycobacterium avium subsp (OASA00348)
Product Page: IL12B antibody (OASA00348)
Antibody Pair Available: OASA00349 Capture OASA00348 Detection
Experiment Type: Cytokine analyses of cell culture supernatants
Bovine IFN-γ was measured by using the Bovigam test kit (Prionics, La Vista, NE). A standard curve was generated using recombinant bovine IFN-γ (3.12 to 100 ng/ml; Thermo Scientific, Rockford, IL). Bovine interleukin-10 (IL-10) was quantified by coating MaxiSorp microtiter plates (Nunc, Rochester, NY) with mouse anti-bovine IL-10 in coating buffer (0.05 M carbonate-bicarbonate buffer, pH 9.6 [MCA2110], 2 μg/ml; Serotec, Raleigh, NC) overnight at room temperature (RT). After washing 5 times with PBS containing 1% Tween 80, samples and dilutions of bovine IL-10 standard (0.3125 to 20 ng/ml) (generous gift from Jayne Hope, Compton, United Kingdom) were added to duplicate wells and incubated at RT for 1 h. After being washed, plates were incubated with mouse anti-bovine IL-10-biotin (MCA2111B; Serotec) for 1 h. Plates were washed again and incubated with avidin-horseradish peroxidase (HRP) conjugate (1:800) (PharMingen, San Diego, CA) for 45 min at RT. After another wash cycle, plates were incubated with substrate solution (40 mM ABTS [2,2′-azino-diethylbenzthiozoline-6-sulfonic acid]; US Biological, Swampscott, MA). Color development was quantified after 30 min by measuring absorbance at 405 nm with a Wallac Victor 1420 multilabel counter enzyme-linked immunosorbent assay (ELISA) plate reader (Perkin-Elmer, Gaithersburg, MD). To quantify bovine IL-4 in culture supernatants, plates were coated with mouse anti-bovine IL-4 (clone CC313, 5 μg/ml; Serotec) in 0.05 M carbonate-bicarbonate buffer and incubated overnight at 4°C. After being washed, plates were blocked with PBS containing 1% bovine serum albumin (BSA) and 0.05% Tween 20 for 1 h at RT. After being washed, samples and dilutions of the standard (recombinant bovine IL-4 [PBP006], 15.6 to 2,000 pg/ml; Serotec) were incubated for 1 h at RT, followed by incubation with mouse anti-bovine IL-4-biotin antibody (clone CC314, 2.5 μg/ml; Serotec) for 1 h at RT, streptavidin-horseradish peroxidase (1:1,000; GE Healthcare, Piscataway, NJ) for 45 min at RT, and then ABTS substrate (US Biological) for 1 h at RT, followed by measuring absorbance at 405 nm. Bovine IL-12 was measured by ELISA after coating plates with mouse anti-bovine IL-12 antibody (MCA1782EL, 8 μg/ml; Serotec) diluted in 0.05 M carbonate-bicarbonate buffer overnight at 4°C. Plates were then blocked with 1% BSA overnight at 4°C, followed by the addition of samples and standard dilutions (55 to 1,500 U/ml; kind gift from Jayne Hope, Compton, United Kingdom), and were incubated for 1 h at RT. This was followed by incubation with biotinylated anti-bovine IL-12 (MCA2173B, 1 μg/ml; Serotec) for 1 h at RT, incubation with streptavidin-horseradish peroxidase (1:800) and ABTS substrate for 1 h at RT, and then measurement of absorbance at 405 nm.
1. There were no discernible effects due to vaccination or in vitro stimulation of cells on the secretion of IL-12 (data not shown). 2. Robust IFN-γ responses were observed in vaccinated calves, but secretion of IL-4, IL-10, and IL-12 was not markedly affected by vaccination.
1: Stabel JR, Waters WR, Bannantine JP, Lyashchenko K. Mediation of host immune responses after immunization of neonatal calves with a heat-killed Mycobacterium avium subsp. paratuberculosis vaccine. Clin Vaccine Immunol. 2011 Dec;18(12):2079-89. Epub 2011 Oct 26. PubMed PMID: 22030370; PubMed Central PMCID: PMC3232711.