Fresh frozen mLNs were cryosectioned and air-dried followed by cold acetone treatment for 10 min at −20°C. Slides were blocked with 10% FBS and 0.1% Tween-20 in PBS for 2 h at room temperature followed by overnight incubation in blocking buffer containing biotinylated PNA (Vector Laboratories) at 4°C. Slides were washed three times with PBS Tween-20 at room temperature before incubation in blocking buffer containing PE-CF594-labeled streptavidin (BD PharMingen), FITC-labeled anti-CD4 (eBioscience) and APC-labeled anti-IgD (eBioscience) for 2 h at room temperature. Slides were washed three times with PBS Tween-20 and PBS at room temperature followed by mounting. The images were captured by a Zeiss LSM 780 confocal microscope and were processed using ZEN 2012 SP1 software (Carl Zeiss).
Pe cf594 labeled streptavidin
Manufactured by BD
PE-CF594-labeled streptavidin is a fluorescently labeled protein that binds to biotin with high affinity. It can be used to detect and localize biotinylated molecules in various biological applications.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 780 confocal microscope, by Zeiss (3 mentions)
Zen 2012 sp1, by Zeiss (1 mentions)
Fitc labeled anti cd4, by Thermo Fisher Scientific (1 mentions)
Biotinylated pna, by Vector Laboratories (1 mentions)
Pefabloc sc, by Merck Group (1 mentions)
N octyl β d glucopyranoside, by Merck Group (1 mentions)
Pe labeled streptavidin, by Thermo Fisher Scientific (1 mentions)
2 protocols using pe cf594 labeled streptavidin
1
Immunofluorescent Staining of Lymph Nodes
2
Multimer Preparation of HLA Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
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DRB1*04:01 (DR0401) and DQB1*03:02 (DQ8) protein was purified from insect cell cultures as previously described (18 (link), 19 (link)). Briefly, DR0401 or DR0401-myc, and DQ0302 protein was purified from supernatants by affinity chromatography, biotinylated with biotin ligase, and dialyzed against phosphate buffer (0.0625 M monobasic sodium phosphate, 0.0375 M monobasic sodium phosphate, pH 6.0). To prepare multimers DR0401, DR0401-myc, or DQ8 monomer was incubated with 0.2 mg/mL peptide, 0.2% n-octyl-β-d-glucopyranoside (Sigma) and 1 mM Pefabloc SC (Sigma) at 37 °C for 72 h. After incubation PE labeled streptavidin (ThermoFisher), PE-CF594 labeled streptavidin (BD), or BV421 labeled streptavidin (BD) was added to conjugate monomers into tetramers. As summarized in Table 1
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