DISCONTINUED Chicken IgY | pre-immune IgY (50 mg)

Product no: AS03 027

AS03 027 | Clonality: Polyclonal | Host: Hen

Replaced by AS10 683 Chicken IgY immunoglobulin fraction (10 mg)

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  • Product Info
  • Host: Chicken
    Clonality: Polyclonal
    Purity: Purified, total IgY (chicken egg yolk immunoglobulin) in PBS pH 8. Contains 0.02 % sodium azide.
    Format: Liquid
    Quantity: 50 mg
    Storage: Store at 4C; make aliquots to avoid working with a stock. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
    Tested applications: Control in immunoassays
    Recommended dilution: The optimal working dilution should be determined by the investigator
  • Additional Information
  • Additional information: Total IgY have been isolated by a PEG precipiation method from a pool of eggs from not immunized hens, IgY has been determined to have a purity over 80 % by SDS PAGE gel electrophoresis
    Additional information (application): Some protocols how to work with IgY antibodies can be found here
  • Background
  • Background:

    Total pre-immune IgY is purified from egg yolks of hens not immunized with any antigens.

  • Product Citations
  • Selected references: Larsson and Voss (2018). Neuroprotective effects of vitamin D on high fat diet- and palmitic acid-induced enteric neuronal loss in mice. BMC Gastroenterol. 2018 Nov 21;18(1):175. doi: 10.1186/s12876-018-0905-9.
  • Protocols
  • How to purify IgY?

    Bulk precipitation methods:

    * ammonium sulfate precipitation (Schwarzkopf and Thiele 1996)

    * dextran sulfate precipitation (Jensenius 1981)

    * polyethylene glycol (PEG) precipitation (Polson and Wechmar 1980)

    A waste disposal should be considered when using methods below:

    * caprylic acid extraction (McLaren et al. 1994)

    * chloroform extraction/PEG combined extraction (Polson 1990)

    * propanol-aceton extraction (Bade and Stegemann 1984)

    Commercially available kits and matrixes (different capacity to process egg yolk):

    * EGGstractTM kit (Promega)

    * EggcellentTM Chicken IgY Purification Kit (Pierce)

    * IMSORB Anti IgY (Biotez)

    * HiTrap® IgY purification (Amersham Pharmacia Biotech)

    Industrial scale methods

    * Column and ultrafiltration method (Kim and Nakai 1996, 1998)

    Recommended Literature

    * This manual contains detailed protocols of IgY purification by different methods as well as trouble shooting.
    "Chicken Egg Yolk Antibodies, Production and Application" R. Schade et al.2000, Springer-Verlag, Lab Manuals, ISBN 3-540-66679-6.

    * "Isolation and Purification of Chicken Egg Yolk immunoglobulins: A review" B. De Meulenaer and A. Huyghebaert 2001, Food and Agricultural Immunology, 13:275-288.

    How towork with IgY antibodies? General recommendations

    IgY present in egg yolk:

    * Western Blot - dilute egg yolk 1:1 or 1:2 with PBS pH 7.4. Centrifuge at room temperature for 30 min. at 4000-6000 x g. Resulting supernatant can be used directly in dilution series like: 1:100, 1: 1000 and 1: 10 000. Testing different dilutions is recommended. Western blot membrane should be washed carefully during whole procedure. Note: Using crude egg yolk in a Western Blot might result in increased background.
    Please, check the protocol: Western Blot using IgY.

    * ELISA - dilute egg yolk several times with PBS + 0.5 % Tween 20. (1: 10 for example)
    Blocking of a plate can be done for 2 hours at room temperature or overnight with 2 % fat free dry milk in PBS + 0.05 % Tween 20 is recommended. Warm up diluted egg yolk to RT before applying on ELISA plate. Incubate egg yolk sample containing primary antibodies for 2 hours at 30ºC or RT.

    * Immunohistochemistry - crude egg yolk IgY is not recommended to use directly, since this might result in a rather high background.
    AgriSera offers IgY antibodies as egg yolk or purified (total IgY fraction) or affinity purified, as requested.

    Purified IgY

    * Western Blot - test IgY concentration from 0.5 - 15 ug/ml of the antibody buffer.
    Please, check the protocol: Western Blot using IgY , Trouble shooting for Western Blot
    * ELISA - prepare the dilution series of antibodies using PBS + 0.5 % Tween 20, proceed as above. Trouble shooting for ELISA
    * Immunohistochemistry - from 1 - 20 ug of IgY/ ml of antibody buffer. Careful optimisation of concentration for each specific antibody is required. IgY can be used in immunohistochemistry after first purification. If obtained results are not satisfactory, antibodies require further
    affinity purification. However, note that after affinity purification some antibodies, which bind very strongly to the antigen might be lost.

    Antibody protocols

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