Donkey anti-Mouse IgG (H&L), DyLight® 488 conjugated

Product no: AS10 1261

AS10 1261 | Clonality: Polyclonal | Host: Donkey | Reactivity: Mouse IgG (H&L)

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  • Product Info
  • Immunogen:

    Purified Mouse IgG, whole molecule

    Host: Donkey
    Clonality: Polyclonal
    Purity: Immunogen affinity purified donkey IgG.
    Format: Lyophilized
    Quantity: 1 mg
    Reconstitution: For reconstitution add 1,1 ml of sterile water, Let it stand 30 minutes at room temperature to dissolve, Prepare fresh working dilutions daily
    Storage: Store lyophilized material at 2-8°C. Product is stable for 4 weeks at 2-8°C after rehydration. For long time storage after reconstitution, dilute the antibody solution with glycerol to a final concentration of 50% glycerol and store as liquid at -20°C, to prevent loss of enzymatic activity. For example, if you have reconstituted 1 mg of antibody in 1,1 ml of sterile water add 1,1 ml of glycerol. Such solution will not freeze in -20°C, If you are using a 1:5000 dilution prior to diluting with glycerol, then you would need to use a 1:2500 dilution after adding glycerol. Prepare working dilution prior to use and then discard. Be sure to mix well but without foaming.
    Recommended dilution: 1 : 20-1 : 2000 for most applications
  • Application Examples
  • Immunofluorescent localization of plant actin in rice protoplasts using anti-plant actin monoclonal recombinant antibodies

    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
  • 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 immunoglobulins

    No reactivity is observed to: non-immunoglobulin mouse serum proteins

  • Background
  • 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.

  • Product Citations
  • Selected references: 38221900
  • Protocols
  • Antibody protocols
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