IL-1 beta Antikörper

Produktnummer ABIN2689746

Produktdetails anti-IL-1 beta Antikörper

Target: IL-1 beta (IL1B) (Interleukin 1, beta (IL1B))

Reaktivität: Human

Wirt: Maus

Klonalität: Monoklonal

Konjugat: Dieser IL-1 beta Antikörper ist unkonjugiert

Applikation: Neutralization (Neut)

Marke: BD Pharmingen™

Produktmerkmale: The AS10 antibody reacts with human interleukin-1β (IL-1β) which is also known as endogenous pyrogen (EP), leukocyte endogenous mediator (LEM), mononuclear cell factor (MCF) and lymphocyte-activating factor (LAF). IL-1β is a proinflammatory cytokine that is synthesized as a precursor of 31 kDa and is converted intracellularly to the mature 17.5 kDa form, after cleavage by the IL-1β-converting enzyme (ICE). In healthy individuals, IL-1β is secreted non-constitutively by blood monocytes, tissue macrophages and dendritic cells. IL-1β is also constitutively expressed in the human hypothalamus. Many malignant tumors express IL-1β as part of their neoplastic nature. The immunogen used to generate the AS10 hybridoma was recombinant human E. Coli-expressed recombinant IL-1β. The AS10 antibody does not cross react with mouse IL-1β. This is a neutralizing antibody. This antibody is routinely tested by immunocytochemistry. Other applications were tested during antibody development only or reported in the literature. PBMC were isolated from human peripheral blood by density gradient centrifugation and were cultured for 6hr at 37 °C with human IFN-γ (20 ng/mL, Cat. No. 554616). The cells were subsequently stimulated with 1 μg/mL LPS (Sigma No. L-8274) an were incubated with GolgiStop™ (Cat.No. 554724) overnight at 37 °C. The activated cells were harvested and the level of IL-1β producing cells was detected by immunocytochemistry using a three-step staining procedure that employs a Biotin Goat Anti-Mouse IgG secondary antibody and a horseradish peroxidase-based detection system. To demonstrate specificity of staining the binding of the AS10 (Cat. No. 550007) antibody was blocked by the preincubation of the purified antibody with excess recombinant human IL-1β protein (Cat. No. 554602, data not shown). (Nomarski optics, original magnification 400 X).

BD Pharmingen™ Purified Mouse Anti-Human IL-1β - Purified - Clone AS10 - Isotype Mouse IgG1, κ - Reactivity Hu - 0.25 mg

Aufreinigung: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Recombinant human IL-1beta

Klon: AS10

Isotyp: IgG1 kappa

Anwendungsinformationen

Applikationshinweise: Optimal working dilution should be determined by the investigator.

Testdurchführung:

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) 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 Pharmingen's IHC Diluent Buffer 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 Cytokine 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 mounding 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.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Konzentration: 0.5 mg/mL

Buffer: Aqueous buffered solution containing ≤0.09 % sodium azide.

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Lagerung: 4 °C

Informationen zur Lagerung: Store undiluted at 4°C.

Details zu IL-1 beta

Target: IL-1 beta (IL1B) (Interleukin 1, beta (IL1B))

Andere Bezeichnung: IL-1beta (IL1B Produkte)

Synonyme: IL-1 antikoerper, IL1-BETA antikoerper, IL1F2 antikoerper, IL-1BETA antikoerper, IL1beta antikoerper, il1-b antikoerper, zgc:111873 antikoerper, IL-1B antikoerper, IL-1beta antikoerper, Il-1b antikoerper, IL1B antikoerper, IL-1 beta antikoerper, IL-1b antikoerper, interleukin 1 beta antikoerper, interleukin 1, beta antikoerper, IL1B antikoerper, il1b antikoerper, Il1b antikoerper

Pathways: NF-kappaB Signalweg, Interferon-gamma Pathway, TLR Signalweg, Negative Regulation of Hormone Secretion, Cellular Response to Molecule of Bacterial Origin, Carbohydrate Homeostasis, Glycosaminoglycan Metabolic Process, Myometrial Relaxation and Contraction, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Autophagie, Cancer Immune Checkpoints, Inflammasome