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Anti il 23p19

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Anti-IL-23p19 is a laboratory reagent that specifically binds to the p19 subunit of the human interleukin-23 (IL-23) cytokine. It is used for the detection and quantification of IL-23 in research applications.

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Lab products found in correlation

Il 23, by R&D Systems (2 mentions) Il 12, by R&D Systems (2 mentions) Propidium iodide, by Thermo Fisher Scientific (1 mentions) Rat igg1κ, by BioLegend (1 mentions) Cytofix cytoperm kit, by BD (1 mentions) Lsrfortessa cell analyzer, by BD (1 mentions) Rmil 12, by Thermo Fisher Scientific (1 mentions) Mog35 55, by Thermo Fisher Scientific (1 mentions)

4 protocols using anti il 23p19

1

Induced EAE Models and Treatments

Cited 1 time
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Active and adoptive transfer EAE model were induced and assessed as previously described49 (link),50 (link). Briefly, for the active EAE, mice were immunized with 200 ng MOG35-55 emulsified in CFA (1:1) subcutaneously followed by intraperitoneal injection of pertussis toxin 200 ng on Day0 and Day2. For the adoptive transfer, donor mice were immunized same as active EAE without pertussis toxin and spleen/draining lymph nodes were harvested 10 days after immunization. The cells were in vitro cultured for 5 days with MOG35-55 (15 μg/ml) under either TH1 cell–polarizing conditions (20 ng/ml IL-12, R&D Systems; 2 μg/ml anti-IL-23p19, eBiosciences) or TH17 cell–polarizing conditions (20 ng/ml IL-23, R&D Systems). For drug treatment, mice were treated with β-glucosylceramide (300 μg per mice) by intravenous on day 10 post the immunization. As for AMP-DNM treatment, mice were intraperitoneally injected with AMP-DNM (1 mg/kg) of ethanol dissolved in PBS daily since the onset of the EAE symptom.
Zhang Q., Liu W., Wang H., Zhou H., Bulek K., Chen X., Zhang C.J., Zhao J., Zhang R., Liu C., Kang Z., Bermel R.A., Dubyak G., Abbott D.W., Xiao T.S., Nagy L.E, & Li X. (2022). TH17 cells promote CNS inflammation by sensing danger signals via Mincle. Nature Communications, 13, 2406.
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2

Induction and Assessment of Active and Passive EAE

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Active EAE was induced and assessed as previously described39 (link). Adoptive transfer (passive) EAE was also induced as previously described40 (link). Briefly, recipient mice were injected with 3.0×107 polarized MOG35−55-specific TH1 or TH17 cells 4 hours after 500 Rads sublethal irradiation. To prepare MOG35−55-specific polarized T cells, donor mice were immunized with MOG35−55 subcutaneously; draining lymph node cells were prepared from donor mice 10 days after immunization. Cells were cultured for 5 days with MOG35−55 at a concentration of 25 μg/ml under either TH1-polarizing conditions (20 ng/ml IL-12, R&D; 2 μg/ml anti-IL-23p19, eBioscience) or TH17-polarizing conditions (20 ng/ml IL-23, R&D). The clinical score was assessed in double-blinded manner. For the sample size, we performed power analysis for the clinical score of EAE, which has an average of 25% coefficient of variance. We determined that with n=15 mice we had 90% power to detect 30% difference between the groups.
Martin B.N., Wang C., Zhang C.J., Kang Z., Gulen M.F., Zepp J.A., Zhao J., Bian G., Do J.S., Min B., Pavicic PG J.R., El-Sanadi C., Fox P.L., Akitsu A., Iwakura Y., Sarkar A., Wewers M.D., Kaiser W.J., Mocarski E.S., Rothenberg M.E., Hise A.G., Dubyak G.R., Ransohoff R.M, & Li X. (2016). T cell-intrinsic ASC critically promotes TH17-mediated experimental autoimmune encephalomyelitis. Nature immunology, 17(5), 583-592.
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3

Single-cell Immune Phenotyping and Cytokine Analysis

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Single-cell suspensions were pretreated with anti-CD16/32 antibody (clone: 2.4G2; TONBO Biosciences, San Diego, CA, USA) to block the Fc receptors. The cells were subsequently stained with the antibody mixture of CD45.2 (clone: 104; BioLegend, San Diego, CA, USA) and CD49f (clone: GoH3; eBioscience, San Diego, CA, USA). Propidium iodide (Invitrogen™, Thermo Fisher Scientific) was used to exclude the dead cells. For intracellular cytokine staining, the cells were stained with surface markers prior to fixation and permeabilization using Cytofix/Cytoperm kit (BD Biosciences, San Jose, CA, USA), according to the manufacturer’s protocol. Anti-IL-23p19 (clone: fc23cpg; eBiosciences) and RatIgG1κ (clone: RTK2071; BioLegend) were used as isotype controls. The stained samples were examined using LSRFortessa™ Cell Analyzer (BD Biosciences). The flow cytometry data were analyzed using FlowJo software (TreeStar, Ashland, OR, USA).
Park Y.J., Kim Y.H., Lee E.S, & Kim Y.C. (2022). Comparative Analysis of Single-Cell Transcriptome Data Reveals a Novel Role of Keratinocyte-Derived IL-23 in Psoriasis. Frontiers in Immunology, 13, 905239.
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4

Experimental Autoimmune Encephalomyelitis Model

Cited 2 times
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For active EAE, mice were immunized subcutaneously (s.c.) on the back with 200 μg of MOG35-55 (MEVGWYRSPFSRVVHLYRNGK) emulsified in CFA (Difco Lab, Detroit, MI) containing 4 mg/ml Mycobacterium tuberculosis H37Ra (Difco). Two hundred ng of pertussis toxin (List Biological Lab, Epsom, England) was given i.p. on days 0 and 2 post immunization (p.i.). For passive EAE, GFAP-shIFN-γR or GFAP-shVec lentivirus injected mice were transferred with 3.0×107 polarized MOG35-55-specific Th1 or Th17 cells/mouse 4 hours after sublethal irradiation (550 Rad). To prepare MOG-specific polarized T cell populations, draining lymph nodes and spleen cells were prepared from mice immunized as described above at day 9 p.i. Cells were cultured for 4 days with MOG35-55 at a concentration of 25 μg/ml under Th1- (20 ng/ml rmIL-12 [PeproTech], 2 μg/ml anti-IL23p19 [eBioscience]) or Th17- (20 ng/ml rmIL-23 [PeproTech]) polarizing conditions (28 (link)). Mice were scored daily for appearance of clinical signs of EAE on a scale from 0 to 5 as described previously (29 (link)): 0, no clinical signs; 1, fully limp tail; 2, paralysis of one hind limb; 3, paralysis of both hind limbs; 4, paralysis of trunk; 5, moribund or death.
Ding X., Yan Y., Li X., Li K., Ciric B., Yang J., Zhang Y., Wu S., Xu H., Chen W., Lovett-Racke A.E., Zhang G.X, & Rostami A. (2015). Silencing IFN-γ binding/signaling in astrocytes vs. microglia leads to opposite effects on CNS autoimmunity. Journal of immunology (Baltimore, Md. : 1950), 194(9), 4251-4264.
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