Rmil 4

Manufactured by BioLegend

Sourced in United States, Japan

RmIL-4 is a recombinant mouse Interleukin-4 (IL-4) protein produced in E. coli. IL-4 is a cytokine that plays a critical role in the regulation of immune and inflammatory responses.

Automatically generated - may contain errors

Lab products found in correlation

9 protocols using rmil 4

Evaluation of Dendritic Cell Maturation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Murine BMDCs were generated as previously described.36 (link) Briefly, bone marrow (BM) cells were cultured in RPMI 1640 with 10 ng/mL rmGM-CSF (E. coli, BioLegend, San Diego, USA) and 5 ng/mL rmIL-4 (E. coli, BioLegend, San Diego, USA), and the medium was replaced on day 3 and 5. Non-adherent cells were harvested on day 7, and CD11c⁺ BMDCs were identified and enriched using FACS (> 90%). The isolated BMDCs were incubated with naked pIL-12, blank Ng(-), and Ng(-)pIL-12 (containing 1 μg/mL pIL-12) for 24 or 36 h, and the surface expression of costimulatory molecules CD40 and CD86, and MHC class II (MHC II) was analysed using flow cytometry (Table 2).Table 2

The Formulations Tested on CT-26 for IL-12 Assay and BMDCs for Maturation in vitro

Acronym	Formulation
Acronym	IL-12 Plasmid (μg/mL)	Chitosan (μg/mL)	γ-PGA (μg/mL)
PBS	-	-	-
pIL-12 (1 μg/mL)	1	-	-
Ng(-)	-	1.6	6.4
Ng(-)pIL-12 (1 μg/mL)	1	1.6	6.4

Zhao H., Wang H., Hu Y., Xu D., Yin C., Han Q, & Zhang J. (2021). Chitosan Nanovaccines as Efficient Carrier Adjuvant System for IL-12 with Enhanced Protection Against HBV. International Journal of Nanomedicine, 16, 4913-4928.

+ Open protocol

+ Expand

Induction of T-cell Subsets from Naive Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Splenocytes from C57BL/6 background Rag1^−/− mice were irradiated with 2,500 Rads (X-rad irradiator) and 3 × 10⁵ cells were seeded into each well of a 96 well plate. Naϊve T cells (CD90.2⁺, CD4⁺, CD62L⁺, CD44⁺, CD25⁻) were sorted from C57BL/6 background WT or Rag1^−/− mice using a FACSaria Fusion cell sorter (BD Biosciences), and 3 × 10⁴ sorted cells were added to each well containing irradiated feeder cells. All cells were cultured with 10 μg/ml anti-CD3 (Biolegend) and 3 μg/ml anti-CD28 (Biolegend) in RPMI with 10% FBS, 1% PSG, and 1x NEAA (Invitrogen), 1x Sodium Pyruvate (Invitrogen), and 0.001% 2-mercaptoethanol. rhIL-2 was used at 30 IU/mL, rmIL-12 and rmIL-4 at 10 ng/mL, rmIL-6 at 20 ng/mL, mTGFβ1 at 1 ng/mL, anti-mIL-4, anti-mIFNγ and anti-mIL-12 at 10 μg/mL (from Biolegend except rhIL-2, rmIL-12, and rmIL-6 from Prepotech). The following cytokines/blocking antibodies were used to skew towards respective Th-subsets: Th0 condition: rhIL-2; Th1 condition: rhIL-2, rmIL-12 and anti-mIL-4; Th2 condition: rhIL-2, rmIL-4, anti-mIFNγ and anti-mIL-12; Th17 condition: rhIL-2, mIL-6, mTGFβ1, anti-mIL-4 and anti-mIL-12; Treg condition: rhIL-2, mTGFβ1, anti-mIL-4 and anti-mIL-12.

Burn T.N., Miot C., Gordon S.M., Culberson E.J., Diamond T., Kreiger P.A., Hayer K.E., Bhattacharyya A., Jones J.M., Bassing C.H, & Behrens E.M. (2022). The RAG1 Ubiquitin Ligase Domain Stimulates Recombination of TCR β and α Genes and Influences Development of αβ T Cell Lineages. Journal of immunology (Baltimore, Md. : 1950).

+ Open protocol

+ Expand

Expanding Self-Renewing Macrophages with Cytokines

Check if the same lab product or an alternative is used in the 5 most similar protocols

Recombinant human (rh) M-CSF (a gift from Morinaga Milk Industry, Kanagawa, Japan), which is cross-reactive to murine cells^{31 (link)}, was used to expand and maintain self-renewing macrophages. Recombinant murine (rm) M-CSF (BioLegend) was also used in a selected experiment. Other cytokines used were as follows: rmGM-CSF (BioLegend), rmIL-4 (BioLegend), rmIL-13 (Peprotech), rmIL-33 (Peprotech), rmIL-34 (BioLegend), and rhRANKL (Oriental Yeast, Tokyo, Japan).

Nasser H., Adhikary P., Abdel-Daim A., Noyori O., Panaampon J., Kariya R., Okada S., Ma W., Baba M., Takizawa H., Yamane M., Niwa H, & Suzu S. (2020). Establishment of bone marrow-derived M-CSF receptor-dependent self-renewing macrophages. Cell Death Discovery, 6, 63.

+ Open protocol

+ Expand

Isolation and Polarization of B Cell Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols

Pan B cells were isolated from spleens of Sox8^−/− or littermate control mice using MojoSort mouse pan B cell isolation kit (BioLegend) in accordance with the manufacturer’s instructions. For IgA⁺ B cell polarization, pan B cells were stimulated with 2 µg/ml F(ab′)2-goat-anti-mIgM (Thermo Fisher Scientific), 5 µg/ml anti-mCD40 (3/23; BioLegend) in complete advanced RPMI 1640 media (Thermo Fisher Scientific) containing 5% vol:vol FCS (MP Biomedicals) supplemented with 1 ng/ml recombinant human (rh) TGF-β1, 5 ng/ml recombinant mouse (rm) IL-5, 20 ng/ml rmIL-21 (all from BioLegend), and 10 nM all-trans retinoic acid (Fujifilm Wako Pure Chemical) for 6 d. For IgG1⁺ and IgE⁺ B cell polarization, pan B cells were stimulated with anti-mIgM and anti-mCD40 in complete media supplemented with 20 ng/ml rmIL-21 and 50 ng/ml rmIL-4 (BioLegend) for 6 d. For IgG2a⁺ B cell polarization, pan B cells were stimulated with anti-mIgM and anti-mCD40 in complete media supplemented with 20 ng/ml rmIL-21 and 50 ng/ml rmIFN-γ (BioLegend) for 6 d.

Kimura S., Kobayashi N., Nakamura Y., Kanaya T., Takahashi D., Fujiki R., Mutoh M., Obata Y., Iwanaga T., Nakagawa T., Kato N., Sato S., Kaisho T., Ohno H, & Hase K. (2019). Sox8 is essential for M cell maturation to accelerate IgA response at the early stage after weaning in mice. The Journal of Experimental Medicine, 216(4), 831-846.

+ Open protocol

+ Expand

Bone Marrow Dendritic Cell Activation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Bone marrow (BM) cells were harvested from femurs and tibiae of C3H/HeN mice, and then cultured in RPMI-1640 medium (Gibco) containing 10% fetal bovine serum (Sigma-Aldrich), 100 IU/mL of Penicillin and 100 μg/mL of Streptomycin (Gibco), in the presence of 25 ng/mL rmGM-CSF and 12.5 ng/mL rmIL-4 (BioLegend) for 8 days at 37°C with 5% CO₂. BMDCs were then stimulated for 4, 8 or 24 hours without or with B. breve, L. rhamnosus or E. coli at a ratio of 1:100.

Li Q., Li Y., Wang Y., Xu L., Guo Y., Wang Y., Wang L, & Guo C. (2021). Oral administration of Bifidobacterium breve promotes antitumor efficacy via dendritic cells-derived interleukin 12. Oncoimmunology, 10(1), 1868122.

+ Open protocol

+ Expand

Murine Allergic Airway Inflammation Model

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mice were anesthetized by isofluorane inhalation, followed by the intranasal administration of rmIL-33 (0.2 µg, Biolegend), rmIL-4 (0.5 µg), rmIL-13 (0.5 µg), rmGM-CSF (0.5 µg), rmIL-5 (0.5 µg), LPS (1 μg, Sigma), CpG (10 μg, Invivogen), Aspergillus protease allergen (0.01U, Sigma), Ragweed pollen extract (300 μg, Greer) or clodronate liposome (C.L.) (30% C.L./PBS, Liposoma B.V.) in 40 µl of PBS. Diphtheria toxin (10 ng/g, Sigma), IL-33R-Fc (10 mg/kg, AstraZeneca), anti-NK1.1 mAb (50 μg, PK136, BioXcell), anti-CCR2 (20 μg, MC-21, provided by Prof. Matthias Mack^{52 (link)}), anti-CCL2 (200 μg, MCP-1, BioXcell), anti-Ly6C/G (200 μg, GR-1, BioXcell), anti-Ly-6G (200 μg, 1A8, BioXcell), anti-CD4 (100 μg, GK1.5, BioXcell), anti-IL-10 (300 μg, JES5-2A5, BioXcell), anti-TGF-β (400 μg, 1D11.16.8, BioXcell), anti-IL-5 (100 μg, TRFK5, BioXcell), rat IgG1, κ (BioXcell), rat IgG2a (BioXcell), or rmIL-33 (0.5 µg, Biolegend) was administered by intraperitoneal injection in 100 µl of PBS. A2AR antagonist (20 μg, SCH 58261, Sigma) was administered in DMSO/PBS (v/v).

Schuijs M.J., Png S., Richard A.C., Tsyben A., Hamm G., Stockis J., Garcia C., Pinaud S., Nicholls A., Ros X.R., Su J., Eldridge M.D., Riedel A., Serrao E.M., Rodewald H.R., Mack M., Shields J.D., Cohen E.S., McKenzie A.N., Goodwin R.J., Brindle K.M., Marioni J.C, & Halim T.Y. (2020). ILC2-driven innate immune checkpoint mechanism antagonizes NK cell anti-metastatic function in the lung. Nature immunology, 21(9), 998-1009.

+ Open protocol

+ Expand

Cytokine-Induced Skin Inflammation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mice were subcutaneously injected each day with the following cytokines dissolved in 100 μl PBS: 500 ng recombinant mouse (rm) IL-33 (BioLegend, 580506) or 500 ng rmIL-13 (BioLegend, 575906) and 500 ng rmIL-4 (BioLegend, 715004) in complex with 2.5 μg anti-mouse IL-4 (Bio X-Cell, 11B11). Whole back skin was collected for all cytokine injections.

Boothby I.C., Kinet M.J., Boda D.P., Kwan E.Y., Clancy S., Cohen J.N., Habrylo I., Lowe M.M., Pauli M., Yates A.E., Chan J.D., Harris H.W., Neuhaus I.M., McCalmont T.H., Molofsky A.B, & Rosenblum M.D. (2021). Early-life inflammation primes a T helper 2 cell–fibroblast niche in skin. Nature, 599(7886), 667-672.

+ Open protocol

+ Expand

Bone Marrow Dendritic Cell Transfer and Treg Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Bone marrow cells were flushed from the tibia and femur of WT BALB/c or PD-L2^KO mice. 2–4 × 10⁶ cells were then cultured in 10 ml cRPMi supplemented with 40 ng/mL Granulocyte maturation colony-stimulating factor (GM-CSF) for 7 to 10 days in a 9-cm diameter tissue culture coated petri dishes. Loosely adherent cells were collected and considered as BMDCs as they were >90% CD11c⁺ based on FACS analysis. In some experiments, WT or PD-L2^KO BMDCs were stimulated overnight with 50 ng/mL rmIL-4 (Biolegend) and the amount of PD-L2 expression was determined by flow cytometry. Cells were washed and resuspended in PBS prior to transferring 1 × 10⁶ BMDCs intravenously (i.v.) into WT Foxp3^GFP or PD-L2^KO Foxp3^GFP recipients. Spleens were then harvested on day 3 and the numbers of CD3⁺CD4⁺CD25⁺Foxp3^GFP+Nrp1^+/− Tregs quantified by flow cytometry.

Hurrell B.P., Helou D.G., Howard E., Painter J.D., Shafiei-Jahani P., Sharpe A.H, & Akbari O. (2022). PD-L2 controls peripherally induced regulatory T cells by maintaining metabolic activity and Foxp3 stability. Nature Communications, 13, 5118.

+ Open protocol

+ Expand

Bone Marrow-Derived Dendritic Cell Activation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Bone marrow was collected from the femurs and tibias of female C57BL/6J mice. Cells cultured in RPMI-1640 medium supplemented with 10% Fetal Bovine Serum (PEAK), L-Glutamine, Na-Pyruvate, Penicillin/Streptomycin, and 2-mercaptoethanol in a 100 mm bacteriological petridish. The cells were then grown for 10 days in the presence of 200 U/mL of rmGM-CSF, with change of media on day 3, 6, and 8. After 10 days non-adherent cells in suspension were harvested and resuspended into RPMI complete with 10 ng/mL rmIL-4 (Biolegend) and 200 U/mL rmGM-CSF (Biolegend), plated at 1 × 10⁶ cells in a 12 well-tissue culture grade plate. One microgram per milliliter of LPS was added per well for LPS maturation of BMDCs. After 18 h cells were harvested stained with cell trace violet dye (Life Technologies) and pulsed with 10 μg/mL of MOG₃₅₋₅₅ in a 24 well-plate for 2 h. Control BMDCs did not receive any MOG₃₅₋₅₅ treatment. CD4 T cells isolated from 2D2 Transgenic mice were stained with CFSE (Life technologies). T cells were plated in a 48 well-tissue culture grade plate along with antigen pulsed BMDCs at a ratio of 10:1 (3 × 10⁶ T cells: 3 × 10⁵ BMDCs). Activation was conducted for indicated time points.

Mitra A., Shanthalingam S., Sherman H.L., Singh K., Canakci M., Torres J.A., Lawlor R., Ran Y., Golde T.E., Miele L., Thayumanavan S., Minter L.M, & Osborne B.A. (2020). CD28 Signaling Drives Notch Ligand Expression on CD4 T Cells. Frontiers in Immunology, 11, 735.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!