All primary murine CD4+ T cells came from homozygous OT-II TCR transgenic mice (Taconic). The following antibodies were used in this study: biotinylated anti–human-CD25 (clone BC96), biotinylated anti-CD3ε (clone 145-2C11), biotinylated anti-CD43 (clone 1B11), and anti-CD28 (clone 37.51) from BioLegend, and anti-CD3ε (clone 145-2C11) from Bio X Cell. Biotinylated I-Ab presenting OVA(323–339) was obtained from the National Institutes of Health Tetramer Facility for pMHC studies. Ovalbumin peptide 323–339 was obtained from AnaSpec, and recombinant human interleukin-2 was obtained from Prometheus. Latrunculin A was obtained from Cayman Chemical, ML-7 from Santa Cruz Biotechnology, Inc., and 2-APB from Abcam.
Ovalbumin peptide 323 339
Manufactured by AnaSpec
Sourced in United States
Ovalbumin peptide 323-339 is a synthetic peptide with the sequence corresponding to amino acids 323-339 of the ovalbumin protein. It is commonly used as a reference standard or control in research applications.
Automatically generated - may contain errors
Lab products found in correlation
Complete freund s adjuvant cfa, by Merck Group (2 mentions)
Lipopolysaccharides (lps), by Merck Group (2 mentions)
Paired antibody kits, by Thermo Fisher Scientific (2 mentions)
2 apb, by Abcam (1 mentions)
Latrunculin a, by Cayman Chemical (1 mentions)
Anti cd3ε clone 145 2c11, by BioXCell (1 mentions)
Anti cd28 clone 37, by BioLegend (1 mentions)
3 protocols using ovalbumin peptide 323 339
1
Murine CD4+ T Cell Activation
2
Antigen-Specific T Cell Activation Assay
3
Antigen-Specific T Cell Activation Assay
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DCs were activated with 100ng/ml LPS (Sigma-Aldrich, MO, USA) overnight and loaded with 1μM ovalbumin peptide 323-339 (AnaSpec, CA, USA) for 90min. After washing, DCs were cultured with purified CD4 OT-II T cells at a ratio of 1:10.
Moloney murine leukaemia virus H19env peptide 123-141 was emulsified in Complete Freund’s Adjuvant (CFA, Sigma-Aldrich, MO, USA) using a sonicator. Mice were immunized with a total of 100μg peptide/CFA subcutaneously into the hind legs. Alternatively, DCs were activated with 100ng/ml LPS (Sigma-Aldrich, MO, USA) overnight and loaded with 50μg/ml H19env peptide for 90min. A total of 1-2×106 LPS-activated peptide-loaded DCs were injected intravenously into mice. DCs were loaded with 25μg/ml Schistosomal egg antigen (SEA, in-house) or 10μg/ml heat-killed Propionibacterium acnes (in-house) overnight. A total of 5×105 cells were injected into the feet of recipient C57Bl/6 mice. 7d post-immunization, popliteal (draining) lymph nodes were extracted, and cells restimulated with antigens (15μg/ml SEA, 1μg/ml heat-killed P. acnes, plate-bound anti-CD3 (clone 2C11)) in vitro for 72hr. Supernatants were taken and cytokine production analyzed by ELISA using paired antibody kits (ebioscience) according to standard protocols.
Moloney murine leukaemia virus H19env peptide 123-141 was emulsified in Complete Freund’s Adjuvant (CFA, Sigma-Aldrich, MO, USA) using a sonicator. Mice were immunized with a total of 100μg peptide/CFA subcutaneously into the hind legs. Alternatively, DCs were activated with 100ng/ml LPS (Sigma-Aldrich, MO, USA) overnight and loaded with 50μg/ml H19env peptide for 90min. A total of 1-2×106 LPS-activated peptide-loaded DCs were injected intravenously into mice. DCs were loaded with 25μg/ml Schistosomal egg antigen (SEA, in-house) or 10μg/ml heat-killed Propionibacterium acnes (in-house) overnight. A total of 5×105 cells were injected into the feet of recipient C57Bl/6 mice. 7d post-immunization, popliteal (draining) lymph nodes were extracted, and cells restimulated with antigens (15μg/ml SEA, 1μg/ml heat-killed P. acnes, plate-bound anti-CD3 (clone 2C11)) in vitro for 72hr. Supernatants were taken and cytokine production analyzed by ELISA using paired antibody kits (ebioscience) according to standard protocols.
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