IL-10 anticorps
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- Antigène Voir toutes IL-10 (IL10) Anticorps
- IL-10 (IL10) (Interleukin 10 (IL10))
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Reactivité
- Humain, Virus
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Hôte
- Rat
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Clonalité
- Monoclonal
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Conjugué
- Cet anticorp IL-10 est non-conjugé
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Application
- Immunocytochemistry (ICC)
- Marque
- BD Pharmingen™
- Attributs du produit
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The JES3-12G8 antibody reacts with human interleukin-10 (IL-10) and viral IL-10. The immunogen used to generate the JES3-12G8 hybridoma was human IL-10. This is a neutralizing antibody. This antibody is routinely tested by immunocytochemical analysis. Other applications were tested during antibody development only or reported in the literature. Human IL-10 Staining. PBMC were isolated from human peripheral blood by density gradient centrifugation and were cultured for 2 days with plate bound anti-human CD3 and soluble anti-human CD28 in the presence of recombinant human IL-2 and recombinant human IL-4. The cells were subsequently harvested, washed and recultured with recombinant human IL-2 and recombinant human IL-4 for an additional 3 days. Finally, the cells were harvested, washed and stimulated with PMA (Sigma) and ionomycin (Sigma) in the presence of GolgiStop™ (Cat. No. 554724) for 4 hours at 37 °C. The activated cells were harvested and the presence of IL-10 producing cells was detected by immunocytochemistry using a three-step staining procedure that employs a biotin goat anti-rat IgG secondary antibody and a horseradish peroxidase-based detection system (see protocol below) (Nomarski optics, original magnification 400 X). To demonstrate specificity of staining the binding of JES3-12G8 (Cat. No. 559076) antibody was blocked by the preincubation of the purified antibody with excess recombinant human IL-10 protein (Cat. No. 554611, data not shown).
BD Pharmingen™ Purified Rat Anti-Human IL-10 - Purified - Clone JES3-12G8 - Isotype Rat IgG2a - Reactivity Hu, Vir - 0.25 mg - Purification
- The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
- Immunogène
- Human IL-10
- Clone
- JES3
- Isotype
- IgG2a
- Top Product
- Discover our top product IL10 Anticorps primaire
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- Indications d'application
- Optimal working dilution should be determined by the investigator.
- Procédure de l'essai
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FOR IMMUNOCYTOCHEMICAL STAINING OF SINGLE-CELL PREPARATIONS This procedure describes the immunoenzymatic technique of staining cytokines within individual cells that are immobilized on microscopic slides via adherence (adherent slides) or centrifugation (cytospins). ADHESION SLIDES 1. Harvest cells and wash them twice in PBS using centrifugation (400 x g for 5 min) to remove residual protein. 2. Adjust the cell concentration at 4-5 x 10e6 cells/mL in PBS. 3. Place 20 μL of the cell suspension in each well of the adhesion slides and let them adhere at room temperature (RT) for 20 min. Please note that the slides should be washed in PBS at RT for 5 min before transferring the cells. 4. Fix cells on slides using fixation buffer for 15 min at RT. 5. Wash slides 2X in PBS with 5 min incubations. 6. Block slides with PBS supplemented with 1 % (w/v) BSA (Sigma, Cat. No. A43-78) for 30 min at RT or 10 min at 37 °C. 7. Wash slides 2X in PBS and proceed with staining or air dry them and store them at -80 °C for future use. 8. Incubate slides with 20 μL of 1 % goat serum and PBS with 0.1 % (w/v) saponin for 30 min at RT. 9. Wash slides 2X with PBS with 5 min incubations. 10. Block endogenous peroxidase activity with Endogenous Peroxidase Blocking Buffer (20 μL/well) for 10 min at RT. 11. Wash 2X in PBS with 5 min incubations. 12. Incubate each well with Avidin (20 μL/well) for 15 min. 13. Wash 2X in PBS with 5 min incubations. 14. Incubate each well with Biotin (20 μL/well) for 15 min. 15. Wash 2X in PBS with 5 min incubations. 16. Incubate each well for 1 hr at RT with 20 μL of purified cytokine-specific antibody or appropriate immunoglobulin isotype control diluted in Antibody Diluent for IHC, Cat. No. 559148, supplemented with saponin 17. Wash slides 2X in PBS with 5 min incubations. 18. Incubate each well with 20 μL of a biotinylated secondary antibody diluted in IHC Diluent Buffer for 30 min at RT. 19. Wash 2X in PBS with 5 min incubations. 20. Apply 20 μL of Streptavidin-HRP (BD Cat. No. 550946) to each well on slides and incubate for 30 min at RT. 21. Wash slides 2X with PBS with 5 minutes incubations. 22. Incubate with DAB Substrate as directed, (BD Cat. No. 550880) for less than 5 min at RT. 23. Stop the development of the color reaction by washing with PBS. 24. The slides are subsequently mounted in short-term storage mounting medium. CYTOSPINS 1. Assemble the Cytospin's sample chamber (e.g. Cytospin 3, Shandon, UK or comparable centrifuge), filter card, slide and cytospin racks according to manufacturer's specifications. 2. Load 40 μL of approximately 1 x 10e6 cells to each sample chamber. 3. Spin slides at 600 rpm for 2 min. 4. Take slides out of the cytospin rack and place them on a staining rack. 5. For fixation and staining please follow the steps 4 through 24 specified above for staining cells on adhesion slides.
- Restrictions
- For Research Use only
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- Concentration
- 0.5 mg/mL
- Buffer
- Aqueous buffered solution containing ≤0.09 % sodium azide.
- Agent conservateur
- Sodium azide
- Précaution d'utilisation
- This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- Stock
- 4 °C
- Stockage commentaire
- Store undiluted at 4°C.
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Epstein-Barr virus transformation induces B lymphocytes to produce human interleukin 10." dans: The Journal of experimental medicine, Vol. 177, Issue 2, pp. 295-304, (1993) (PubMed).
: "Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples." dans: Immunological reviews, Vol. 127, Issue 9-10, pp. 5-24, (1992) (PubMed).
: "Presence of interleukin 10 (IL-10) in the ascites of patients with ovarian and other intra-abdominal cancers." dans: Cytokine, Vol. 4, Issue 5, pp. 385-90, (1992) (PubMed).
: "IL-10 is produced by subsets of human CD4+ T cell clones and peripheral blood T cells." dans: Journal of immunology (Baltimore, Md. : 1950), Vol. 149, Issue 7, pp. 2378-84, (1992) (PubMed).
: "Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures." dans: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, Vol. 29, Issue 4, pp. 577-80, (1981) (PubMed).
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Epstein-Barr virus transformation induces B lymphocytes to produce human interleukin 10." dans: The Journal of experimental medicine, Vol. 177, Issue 2, pp. 295-304, (1993) (PubMed).
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- Antigène
- IL-10 (IL10) (Interleukin 10 (IL10))
- Autre désignation
- IL-10 (IL10 Produits)
- Synonymes
- anticorps CSIF, anticorps GVHDS, anticorps IL-10, anticorps IL10A, anticorps TGIF, anticorps Il-10, anticorps IL10X, anticorps interleukin 10, anticorps IL10, anticorps Il10
- Pathways
- Cellular Response to Molecule of Bacterial Origin, Regulation of Leukocyte Mediated Immunity, Production of Molecular Mediator of Immune Response, Maintenance of Protein Location, Cancer Immune Checkpoints
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