Rabbit anti-Chicken IgY (H&L), HRP conjugated

Product no: AS10 1489
AS10 1489 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Chicken IgG/Y (H&L)
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  • Product Info
  • Immunogen: Purified Chicken IgY (H&L), whole molecule
    Host: Rabbit
    Clonality: Polyclonal
    Purity: Immunogen affinity purified rabbit 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, Centrifuge to remove any particulates, Prepare fresh working dilutions daily
    Storage: Store lyophilized material at 2-8°C.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.
    Tested applications: ELISA (ELISA), Immunohistochemistry (IHC), Western blot (WB)
    Recommended dilution: The optimal working dilution should be determined by the investigator
  • Reactivity
  • Confirmed reactivity: Heavy chains on chicken IgY and light chains on all chicken immunoglobulins,
    Not reactive in: Non-immunoglobulin chicken serum proteins
  • Application Examples
  • Application example

    western blot with chicken anti-RbcL antibodies

    1 µg of total protein from samples such as Arabidopsis thaliana leaf (1)Hordeum vulgare leaf (2), Zea mays leaf (3)Chlamydomonas reinhardtii total cell (4), were extracted with Protein Extraction Buffer PEB (AS08 300). Samples were diluted with 1X sample buffer (NuPAGE LDS sample buffer (Invitrogen) supplemented with 50 mM DTT and heat at 70°C for 5 min and keept on ice before loading. Protein samples were separated on 4-12% Bolt Plus gels,  LDS-PAGE and blotted for 70 minutes to PVDF using tank transfer. Blots were blocked immediately following transfer in 2% blocking reagent (GE RPN 2125; Healthcare) or 5% non-fat milk dissolved in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 10 000 (in blocking reagent) for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, and then washed 1x15 min and 3x5 min with TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, secondary antibody AS10 1489, Agrisera) diluted to 1:25 000 in blocking reagent for 1h at room temperature with agitation. The blots were washed as above. The blot was developed for 5 min with chemiluminescent detection reagent according the manufacturers instructions. Images of the blots were obtained using a CCD imager (VersaDoc MP 4000) and Quantity One software (Bio-Rad). Exposure time was 30 seconds.
  • Additional Information
  • Additional information:

    Purity of this preparation is > 95% based on SDS-PAGE. Antibody concentration is 1.0 mg/ml. Antibody is supplied in 10 mM sodium phosphate, 0.15 M sodium chloride, pH 7.2.1 % (w/v) B, Protease/IgG free. Contains 0.1 % (v/v) Kathon CG as preservative of bacterial growth.

  • Background
  • Background: Rabbit anti-chicken IgY (H&L) is a secondary antibody conjugated to HRP, which binds to in immunological assays. Antibody is affinity purified using solid phase chicken IgY.
    Chicken egg yolk immunoglobulin (IgY) has been also called chicken IgG.
  • Product Citations
  • Selected references: Li et al. (2022), The effects of Ni availability on H2 production and N2 fixation in a model unicellular diazotroph: The expression of hydrogenase and nitrogenase. Limnol Oceanogr, 67: 1566-1576. https://doi.org/10.1002/lno.12151
    Panayiotou et al. (2018). Viperin restricts Zika virus and tick-borne encephalitis virus replication by targeting NS3 for proteasomal degradation.J Virol. 2018 Jan 10. pii: JVI.02054-17. doi: 10.1128/JVI.02054-17.
  • Protocols
  • Antibody protocols
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