Donkey anti-Mouse IgG (H&L), DyLightŪ 488 conjugated
AS10 1261 | Clonality: Polyclonal | Host: Donkey | Reactivity: Mouse IgG (H&L)
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Purified Mouse IgG, whole molecule
Immunofluorescent localization of actin on suspension culture of Oryza sativa ssp. japonica cv. 'Unggi 9', using anti actin (AS21 4615) and anti-mouse IgG DyLight® conjugated secondary antibodies (AS10 1261). Few representative actin filaments are highlighted by yellow arrowheads. DAPI staining of nuclei is pseudocolored red.
Material: Suspension cultures of Oryza sativa ssp. japonica cv. 'Unggi 9'
Fixation: Packed cell volume to fixer ratio: 250 µl : 5ml Fixer composition and buffer: 4% (w/v) paraformaldehyde (freshly prepared as 8% stock and 0.2 µm filtered) in Phosphate Buffered Saline (PBS), pH 7.4 (2x stock, 0.2 µm filtered)
Container and method: in 6 cm Petri dish, gentle shaking at room temperature (RT)
Duration: 40 minutes. Triton X100 is not used in fixer. Cells were not shaken during the first 5 mins of fixation to allowed to partially recover from osmotic shock induced by formaldehyde.
Cell wall digestion: Yes Packed cell volume to enzyme ratio: 100ul : 2ml Enzyme composition: 1% (A) 1.2% (R) Cellulase (chromatically purified, powder, Worthington) 1% (A) 1.2% (R) Pectinase (protease free, liquid, Sigma) Buffer: 0.5% (w/v) MES buffer, pH 5.6
Container and method: in 2 ml microfuge tube by rolling at room temperature (RT)
Duration: 30 mins (A) or 60 mins (R)
Membrane permeabilization: Triton-X100 (0.35%), 7 min/RT
Antigen retrieval: no
Blocking buffer: Fish gelatin (5% v/v)
Washing buffer: PBS
Primary antibody dilution and incubation time: 1:500, 1hr/RT
Secondary antibody dilution and incubation time and supplier: DyLight® 488 (AS10 1261) 1:600, 45 min/RT
Co-staining of the nucleus (DAPI): Yes
Cell wall and nucleus staining: 100 ng/ml DAPI
Courtesy of Dr. Ferhan Ayaydin, Hungarian Centre of Excellence for Molecular Medicine (HCEMM), Szeged, Hungary.
Conjugate is present in 10 mM Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 1 % (w/v) BSA, Protease/IgG free. 0.05 % (w/v) sodium azide is added as preservative.Based on immunoelectrophoresis, this antibody reacts with: heavy (γ) chains on mouse IgG, light chains on all mouse immunoglobulins
No reactivity is observed to: non-immunoglobulin mouse serum proteins
Donkey anti-Mouse IgG (H&L), DyLight®488 Conjugated is a secondary antibody conjugated to DyLight® 488, which binds to Mouse IgG (H&L) in immunological assays.
DyLight® 488 has Amax = 493 nm, Emax = 518 nm. Antibodies are purified using solid phase Mouse IgG (H&L)
DyLight® is a registered trade mark of Thermofisher Inc., and its subsidaries.