IL-12 / IL-23 Antibody | CC301

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IL-12 / IL-23 Antibody | CC301 gallery image 1

Published customer image:
IFN-gamma response (log10 scale) in mesenteric lymph nodes, Peyer’s Patches, popliteal LN and PBMC against GRA7, TLA and MIC3 in function of time.

From: Verhelst D, De Craeye S, Entrican G, Dorny P, Cox E. Parasite distribution and associated immune response during the acute phase of Toxoplasma gondii infection in sheep. BMC Vet Res. 2014 Dec 16;10(1):293.

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IL-12 / IL-23 Antibody | CC301 gallery image 2

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Comparative IL-12 and IFN&gamma responses of neonatal and adult MLN cells following TLR agonist stimulation. Neonatal (closed circles) and adult (open squares) MLN cells were cultured in vitro with or without 12.5 µg/ml polyI:C, 0.5 µg/ml R-848 or 5 µg/ml Gardiquimod. Supernatants were harvested after 48 h of culture and ELISA was carried out for IL-12 (A) and IFN? (B) secretion. Non-specific cell stimulation was also carried out with 50 ng/ml PMA combined with 500 ng/ml ionomycin, with the supernatants assayed for IFN? detection 48 h later (C). Each circle or square represents one neonate or one adult, respectively. Medians are shown for each stimulus. Non-parametric Mann-Whitney tests were used to compare data for neonates and adults: *p=0.01; **p=0.005; ***p=0.001; ****p=0.0005.

From: Ferret-Bernard S, Remot A, Lacroix-Lamandé S, Metton C, Bernardet N, et al. (2010) Cellular and Molecular Mechanisms Underlying the Strong Neonatal IL-12 Response of Lamb Mesenteric Lymph Node Cells to R-848. PLoS ONE 5(10): e13705.

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IL-12 / IL-23 Antibody | CC301 gallery image 3

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MLN and spleen cell IL-12 responses to R-848 stimulation as a function of age. MLN and spleen cells from 6- to 14-day-old neonates (closed circles), 20-day-old lambs (closed diamonds) and adults (open squares) were cultured in vitro for 48 h, with or without 0.5 µg/ml R-848. At the end of the culture period, supernatants were harvested and ELISA carried out to assess IL-12 secretion by MLN cells (A) or spleen cells (B). Medians are shown for each stimulus. Non-parametric Mann-Whitney tests were used to compare data for neonates, 20-day-old lambs and adults: *p=0.01; **p=0.005; ***p=0.001; ****p=0.0005.

From: Ferret-Bernard S, Remot A, Lacroix-Lamandé S, Metton C, Bernardet N, et al. (2010) Cellular and Molecular Mechanisms Underlying the Strong Neonatal IL-12 Response of Lamb Mesenteric Lymph Node Cells to R-848. PLoS ONE 5(10): e13705.

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IL-12 / IL-23 Antibody | CC301 gallery image 4

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Roles of TGFß1 and IL-10 in regulating IL-12 responses to R-848. MLN cells from neonates (closed circles) and adults (open squares) were cultured in vitro for 48 h, with or without 0.5 µg/ml R-848, in the presence of rhTGFß1 (n = 7) (A) or rovIL-10 (n = 5) (B). Supernatants were harvested and ELISA was carried out to assess IL-12 secretion. The mean ± SEM level of IL-12 secretion is shown (A, B). RNA was extracted and purified from freshly isolated MLN cells. IL-10 mRNA levels were determined by quantitative RT-PCR. Median normalised values are presented for neonates (closed circles) and adults (open squares) (C). MLN cells from neonates (closed circles) and adults (open squares) were stimulated in vitro for 48 h with or without 0.5 µg/ml R-848. Supernatants were harvested and ELISA carried out to assess IL-10 secretion. Medians are indicated. Non-parametric Mann-Whitney tests were used to compare data for neonates and adults: **p=0.001 (D). RNA was extracted and purified from freshly isolated MLN cells. Foxp3 mRNA levels were determined by quantitative RT-PCR. Median normalised values are presented for neonates (closed circles) and adults (open squares) (E). Comparison of the IL-12 response of neonate MLN cells to R-848 stimulation in culture medium supplemented with 10%FCS, 10% neonate autologous plasma or 10% adult plasma. Paired t-test between neonate and adult plasma were non-significant (F).

From: Ferret-Bernard S, Remot A, Lacroix-Lamandé S, Metton C, Bernardet N, et al. (2010) Cellular and Molecular Mechanisms Underlying the Strong Neonatal IL-12 Response of Lamb Mesenteric Lymph Node Cells to R-848. PLoS ONE 5(10): e13705.

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IL-12 / IL-23 Antibody | CC301 gallery image 5

Published customer image:
Main cell population producing IL-12 in response to R-848. Neonatal MLN cells were cultured in vitro with or without 0.5 µg/ml R-848 and, at various time points, supernatants were harvested and ELISA was carried out for IL-12. Mean ± SEM IL-12 secretion is shown for each time point (A). Neonatal MLN cells were cultured in vitro with (black bars) or without (hatched bars) 0.5 µg/ml R-848 for 8 h, with brefeldin A added for the last 5 h. Cells were harvested for IL-12 intracellular staining combined with labelling of CD11b, CD14, MHC class II and CD205. Three independent experiments were carried out and the data shown are the mean proportions of IL-12+ cells after gating on CD11b+, CD14+, MHC class II+ or CD205+ MLN cells (B).

From: Ferret-Bernard S, Remot A, Lacroix-Lamandé S, Metton C, Bernardet N, et al. (2010) Cellular and Molecular Mechanisms Underlying the Strong Neonatal IL-12 Response of Lamb Mesenteric Lymph Node Cells to R-848. PLoS ONE 5(10): e13705.

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  • Product Type
    Monoclonal Antibody
  • Clone
    CC301
  • Isotype
    IgG2a
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    MCA1782ELE, ES, F *, FN, WBdatasheet pdfdatasheet pdf0.5 mg
    MCA1782EL
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • Mouse anti Bovine Interleukin-12/23 antibody, clone CC301 recognizes the p40 subunit of bovine interleukin-12, and binds to both the free subunit and the intact heterodimer. The p40 subunit is also known as IL-12B and can form a heterodimer with either IL-12A or IL-23A.

      This antibody may be used as a capture antibody in an ELISA in combination with biotinylated clone CC326 as a detection reagent.
    • Intended Use
    • Target Species
      Bovine
    • Species Cross-Reactivity
      Target SpeciesCross Reactivity
      Humanyes
      Sheepyes
      Mouseno
      African Buffaloyes
      N.B. Antibody reactivity and working conditions may vary between species.
    • Product Form
      Purified IgG - liquid
    • Reconstitution
    • Preparation
      Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
    • Preservative Stabilisers
      None present
    • Immunogen
      Recombinant bovine IL-12.
    • Purity
    • Approx. Protein Concentrations
      IgG concentration 1.0 mg/ml
    • Reagents In The Kit
    • Preparing The Antibody
    • Endotoxin Level
      <0.01 EU/ug
    • Test Principle
    • Buffer Solution
      Phosphate buffered saline
    • Fusion Partners
      Spleen cells from immunised BALB/c mice were fused with cells of the mouse SP2/0 myeloma cell line.
    • Storage
      Store at -20oC only.

      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.
    • Shelf Life
      18 months from date of despatch.
    • GO Terms
      growth factor activity
      natural killer cell activation
      interferon-gamma biosynthetic process
      cytokine receptor activity
      membrane
      positive regulation of activated T cell proliferation
      T-helper cell differentiation
      cytokine activity
      interleukin-12 receptor binding
    • UniProt
    • Entrez Gene
    • Acknowledgements
    • Regulatory
      For research purposes only
    • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

    • Application NameYesNoMin DilutionMax Dilution
      ELISA8ug/ml
      ELISpot
      Functional Assays1/100
      Western Blotting1/501/100
      Flow Cytometry(1)1/10
      (1)
      Membrane permeabilisation is required for this application. Bio-Rad recommends the use of Leucoperm (Product Code BUF09) for this purpose.

    • Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.
    • Technical Advice
    • Recommended Protocol
    • Flow Cytometry
      Use 10ul of the suggested working dilution to label 106 cells in 100ul.
    • ELISA
      This antibody may be used as a capture antibody in a sandwich ELISA in combination with biotinylated clone CC326 (MCA2173B) as detection reagent, see Bannerman, D.D.et al.
    • Immunohistology
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Western Blotting
    • Instructions For Use

    Additional IL-12 / IL-23 Antibody Formats

    Formats Clone Applications Sizes available
    IL-12 / IL-23 Antibody : Low Endotoxin CC301 E, ES, F *, FN, WB 0.5 mg
    • Copyright © 2016 Bio-Rad

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      Recommended Negative Isotype Control

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        Useful Reagents

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              References

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