Ova peptide 323 339

Manufactured by InvivoGen

OVA-peptide 323–339 is a synthetic peptide derived from the chicken ovalbumin protein sequence. It consists of the amino acid residues 323 to 339. The peptide is commonly used in research applications involving cell-based assays and immunological studies.

Automatically generated - may contain errors

Lab products found in correlation

Celltrace violet, by Thermo Fisher Scientific (2 mentions) E coli lps, by Merck Group (1 mentions) Pan t isolation kit, by Miltenyi Biotec (1 mentions) Interleukin 2 (il 2), by BioLegend (1 mentions) Tgf β, by Thermo Fisher Scientific (1 mentions) Dynabeads mouse t activator cd3 cd28, by Thermo Fisher Scientific (1 mentions) Rpmi 1640 growth medium, by Thermo Fisher Scientific (1 mentions) 3h thymidine, by MP Biomedicals (1 mentions) Siinfekl, by InvivoGen (1 mentions) Dq ova, by Thermo Fisher Scientific (1 mentions) Easysep mouse cd4 t cell isolation kit, by STEMCELL (1 mentions) Lipopolysaccharides (lps), by Merck Group (1 mentions) αcd28, by BioLegend (1 mentions) αcd3 145 2c11, by BioLegend (1 mentions) α cd23, by BioLegend (1 mentions)

Spelling variants of this product

OVA peptide (9 citations)

5 protocols using ova peptide 323 339

Quantifying B-T Cell Interactions

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

Naïve B cells from each indicated group of mice were purified
with mouse anti-CD43 (Ly-48) microbeads, stained with 3 µM Cell Trace
Violet (Invitrogen, Green Island, NY) and activated with 25 µg/ml
soluble anti-IgM (Jackson Immunoresearch) and 1 µg/ml LPS (E.coli LPS,
Sigma Aldrich) for 24h h at 37°C in a CO₂ incubator.
Simultaneously, naïve T cells from OT-II transgenic mice were purified
with a pan-T isolation kit (Miltenyi Biotech), stained with 3 µM CFSE,
and activated with 10 µg/ml anti-CD3 (clone 145-2C11, UCSF monoclonal Ab
core) and 2 µg/ml anti-CD28 (clone PV-1, UCSF monoclonal Ab core) for
24h. For the last 6 h of stimulation, 10 µg/ml OVA peptide
(323–339) (InvivoGen, San Diego, CA) was added to the B cells.
Thereafter, B and T cells were washed. 1×10⁶ B cells and
1×10⁶ T cells were mixed together in a total volume of
100 µl of FACS buffer and briefly centrifuged at 4 °C for 1 min
at 1500 rpm. B:T mixtures were then immediately fixed (time-0 min) or incubated
in a 37 °C water bath for the indicated time points (2, 5, 15, 30 and 60
min). Interacting B and T cells were co-cross-linked by the addition of 100
µl of 4% paraformaldehyde at the end of each time point. Samples
were acquired by flow cytometry.

Soni C., Domeier P.P., Wong E.B., S.h.w.e.t.a.n.k., Khan T.N., Elias M.J., Schell S.L., Lukacher A.E., Cooper T.K, & Rahman Z.S. (2015). Distinct and synergistic roles of FcγRIIB deficiency and 129 strain-derived SLAM family proteins in the development of spontaneous germinal centers and autoimmunity. Journal of autoimmunity, 63, 31-46.

+ Open protocol

+ Expand

Suppression and Induction of Regulatory T Cells

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

For the suppression assay, 2.5 × 10⁴ CD4⁺CD44⁻CD62L⁺ naive T cells were isolated from the spleens of BALB/c mice and co-cultured with live Foxp3^GFP or PD-L2^KO Foxp3^GFP sorted splenic pTregs (CD3⁺CD4⁺CD25⁺Foxp3^GFP+Nrp1⁻) at 1:2: 1:4, 1:8, and 1:16 Treg:T cell ratios in cRPMi as described above for 72 h. For the BMDC co-cultures, WT or PD-L2^KO BMDCs were generated as described above. 4 × 10⁵ naive KJ126⁺CD4⁺CD25⁺CD62L⁺ splenic T cells were then isolated and co-cultured with WT or PD-L2^KO BMDCs at a 1:20 DC to T-cell ratio. Cells were cultured in cRPMi containing 20 ng/mL IL-2 (Biolegend) and 5 ng/mL TGF-β (ebioscience), 100 ng/ml OVA-peptide 323–339 (Invivogen) and T cells were stimulated with a 1:1 ratio of Dynabeads Mouse T-activator CD3/CD28 (Gibco) for 72 h.

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

Measurement of Antigen-Specific T Cell Proliferation

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

To assess cell proliferation, mice were euthanized on day 7 and 1 × 10⁶ splenocytes were cultured with 0–250 μg/ml OVA-peptide 323–339 (Invivogen) for 3 or 4 days in cRPMi. cRPMi was prepared using RPMI 1640 growth medium (Gibco), 10% Fetal Bovine Serum (Promega scientific) and 100 U/ml Penicillin/Streptomycin. Prior to culture, splenocytes were labeled with 1 µM CellTrace Violet (CTV, Thermofisher) for 20 min and stained on day 4 with CD3 and CD4 to isolate proliferating T cells and measure CTV dilution. In some experiments, proliferation was measured following 3 days of culture, where 1μ Ci ³H thymidine (MP Biomedicals, Solon, OH) was then added overnight and the amount of thymidine incorporation was quantified from harvested cells with a scintillation counter (Beckton Dickinson).

+ Open protocol

+ Expand

Immunological Analysis of Lymphocytes Post-Ear Immunization

Cited 1 time

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

Mice were sacrificed at 7 or 28 days post-immunization in the ear, to collect SDLN and skin. SDLN were macerated and filtered, and skin-infiltrating lymphocytes were obtained as previously described (28 (link)). Briefly: Skin cell suspensions were obtained by enzymatic digestion with Liberase TL and DNAse, then chopped and incubated under the same conditions. Next, enzymatic digestion was stopped, and cell suspensions were filtered followed by the addition of DNAse. Finally, cells were washed with PBS, counted and stained. SDLN cells were re-stimulated as previously reported (28 (link)). Briefly: Cells were incubated for 48 h with 1 mg/ml of SIINFEKL (InvivoGen) and OVA peptide 323–339 (InvivoGen), followed by cell stimulation cocktail plus protein transport inhibitor for 4 h at 37°C. Then the lymphocytes were collected, washed, and stained.

León-Letelier R.A., Castro-Medina D.I., Badillo-Godinez O., Tepale-Segura A., Huanosta-Murillo E., Aguilar-Flores C., De León-Rodríguez S.G., Mantilla A., Fuentes-Pananá E.M., López-Macías C, & Bonifaz L.C. (2020). Induction of Progenitor Exhausted Tissue-Resident Memory CD8+ T Cells Upon Salmonella Typhi Porins Adjuvant Immunization Correlates With Melanoma Control and Anti-PD-1 Immunotherapy Cooperation. Frontiers in Immunology, 11, 583382.

+ Open protocol

+ Expand

Isolation and Activation of Murine T-cells and Dendritic Cells

Cited 1 time

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

Total leukocytes were isolated from LN and spleen from 8 to 12 week old mice as previously described [85 (link)]. CD4+ T-cells were isolated from the pooled cell suspensions using an EasySep™ Mouse CD4+ T Cell Isolation Kit from Stemcell Technologies (Vancouver, Canada) following the manufacturer’s instructions. Foxp3+ Treg induction protocols were as previously described [52 (link)]. Polyclonal T-cell activation was performed as previously described [87 (link)], pre-coating plates with 10 μg/ml αCD3 (145–2C11, Biolegend) in PBS overnight at 4°C. Coated plates were washed and cells were plated with 1 μg/ml soluble αCD28 antibody (37.51, Biolegend). Bone marrow-derived dendritic cells (BMDC) were generated as previously described [86 (link)]. DCs were activated overnight with 1 ug/ml LPS (Sigma) along with OVA peptide 323–339 (Invivogen) for antigen specific activation. For polyclonal activation via APC, BMDC Fc receptors were pre-loaded with 2.5 ug/ml αCD23 (Biolegend) and .5 ug/ml αCD28 (Biolegend) for two hours before culturing with target CD4+ T-cells. Antigen processing assays were performed according to manufacturer’s protocol for DQ-OVA (Thermo Fisher Scientific). A phagocytosis assay using these cells in conjunction with pHrodo-labelled particles was performed as previously described [86 (link)].

Marshall P.L., Nagy N., Kaber G., Barlow G.L., Ramesh A., Xie B.J., Linde M.H., Haddock N.L., Lester C.A., Tran Q.L., de Vries C., Hargil A., Malkovskiy A., Gurevich I., Martinez H.A., Kuipers H.F., Yadava K., Zhang X., Evanko S.P., Gebe J.A., Wang X., Vernon R.B., de la Motte C., Wight T.N., Engleman E.G., Krams S.M., Meyer E, & Bollyky P.L. (2020). Hyaluronan synthesis inhibition impairs antigen presentation and delays transplantation rejection. Matrix biology : journal of the International Society for Matrix Biology, 96, 69-86.

+ 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!