Donkey anti-Mouse IgG (H&L), DyLightŪ 488 conjugated
AS10 1261 | Clonality: Polyclonal | Host: Donkey | Reactivity: Mouse IgG (H&L)

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Product Information
Purified Mouse IgG, whole molecule
Reactivity
Application examples

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.
Hydrophilization: no
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.
Additional information
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 immunoglobulinsNo reactivity is observed to: non-immunoglobulin mouse serum proteins
Background
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.