Standing out in the field of IgY Immunotechnology

Click here for the IgY Purification Record printout.

Name of Operator: ________________________  Date: _____________________

Chicken No.: _______  Immunizing Antigen: ______________ Lot size (# eggs): _______

Date of Eggs: _______________________Yield IgY per ml egg yolk (mg) __________

IgY Concentration (mg/ml): ________  Total IgY Yield: ____________ Total vol. (ml) _______

Instructions

  1. Assume that 1 gram of yolk is equal to 1 ml.
    Volume of egg yolks = _________ ml (vol. A)

  2. Add 5 times the egg yolk volume (A) of cold Reagent A to the container and stir gently:
    Volume of Reagent A = 5 x (A) = _________ ml   Lot no. Reagent A: __________________

  3. Place in fridge (at least 2 hours — up to overnight):
    Incubation Time = _________ hr

  4. Centrifuge solution at 2,000 x g ( 4°C) for 15 min.  Collect supernatant into graduated cylinder and discard lipidy pellet. by filtering through gauze inserted into filter paper. The supernatant should be colourless and translucent. If particulates are present, filter through cotton gauze or filter paper. Measure volume of supernatant.
    Volume of supernatant (ml) = _______ (vol. B)     Colourless, translucent supt? ______ yes ______ no

  5. To supernatant (vol. B), add the same volume of cold Reagent B and mix gently. Let sit for at least 1 hr in the fridge (up to overnight).
    Volume Reagent B added (vol. C): ________ ml    Lot no. Reagent B: ___________________  Incubation time: ________ hr

  6. Centrifuge 4°C at 2,000 x g for 15 min. Resuspend the pellet in the original egg yolk volume (vol. A) with PBS:
    Final IgY volume (vol. A): _____ ml

  7. To determine the IgY conc. and yield, prepare two 1/10 dilutions of the purified IgY in PBS (ie add 100 ul of IgY to 900 ul PBS). Measure the absorbance at 280 nm using PBS as blank.
    Abs#1= _______ Abs#2= _________   Avg. Abs = ________
    Concentration = (Avg Abs x 10)/1.35 = ________ mg/ml
    Yield = Concentration x Final Volume (vol. A) = ________ mg

  8. Filter sterilize or add a preservative and refrigerate. IgY activity will remain unchanged for at least one year.
    Filter sterilized? ____ yes _____ no
    Preservative? ____ yes _____ no
    If yes, what kind? _____________________    Concentration: _______________

    Click here for the IgY Purification Record printout.

Kit and Kit Components Costs

Code:
Product:
Price:
IK 500
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IK 2000
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IK 4000
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IK 8000
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RA500
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RA2000
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RA4000
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RA8000
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RB500
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RB2000
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RB4000
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RB8000
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ES001
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Our Guarantee

We guarantee that if you follow the Kit Instructions and if you do not obtain at least 4 mg of 90% pure IgY per ml yolk, we will refund the cost of the kit.

Other products you may require:
Normal Non-Immune IgY
Anti-Chicken Immunoglobulin Secondary Reagents

Related information:
Kit Instructions
Immunization Protocol for Chickens
IgY Purification Record

Don't want to purify your IgY yourself?
Ask us to perform your chicken IgY purification for you >> Antibody purification

The answer is simple.

Compared to the competition, not only is our chicken antibody purification kit cheaper to buy (per mg IgY purified) but the IgY yield obtained per gram egg yolk is, on average, 50% higher.

Unlike other IgY purification kits, our kit is polyethylene glycol (PEG) - free. High molecular weight PEG may interfere with your antibody assay and is difficult to remove.

Our antibody purification kit comes with detailed instructions and technical support is always available. This email address is being protected from spambots. You need JavaScript enabled to view it. with any questions you may have.

Kit Contents

5 egg yolk isolations
(Cat. No. IK 500)
20 egg yolk isolations
(Cat. No. IK 2000)
80 egg yolk isolations
(Cat. No. IK 8000)
5 egg yolk isolations
(Cat. No. IK 500)
  • Reagent A (500 ml)
  • Reagent B (500 ml)
  • Egg Separator
  • Instructions
20 egg yolk isolations
(Cat. No. IK 2000)
  • Reagent A (4 x 500 ml)
  • Reagent B (4 x 500 ml)
  • Egg Separator
  • Instructions
80 egg yolk isolations
(Cat. No. IK 8000)
  • Reagent A (2 x 3.8 Litres)
  • Reagent B (2 x 3.8 Litres)
  • Egg Separator
  • Instructions

Kit Storage Conditions

  • Keep at room temperature until ready to use.
  • Reagents A and B must be refrigerated before use.

What you will need

  • Egg yolks
  • Distilled Water
  • Phosphate-buffered saline (PBS)
  • Pasteur pipette
  • Balance
  • Beaker, graduated cylinder
  • Stir plate and stir bar
  • Refrigerator
  • Centrifuge

Separate Kit Components Available

Perhaps a 90% pure IgY preparation is not necessary for your experimental procedure. Delipidation of the egg yolk provides the researcher with a diluted IgY solution (also containing other yolk proteins) which, concentrated and dialyzed, can be used for many applications. To delipidate your egg yolk only, egg separators and Reagent A are available separately (see here).

Reagent B, our IgY precipitation reagent, is also available separately to purify IgY from other yolk proteins after delipidation. 

  1. instructionsIt is important that Reagents A and B and your eggs are at 4°C before use!
  2. Separate cold egg yolk from the white using the egg separator. Rinse yolk with distilled water and roll on paper towel to remove adhering egg white. Puncture yolk membrane with Pasteur pipette and allow yolk to drain into tared beaker. Note the weight of the yolk. Assume that 1 ml of yolk is equal to 1 gram.
  3. Add 5 volumes of cold Reagent A to the yolks very slowly while stirring gently and continuously until well mixed. Avoid foaming.
  4. Allow diluted yolk to stand for at least 2 hours or up to 24 hours at 4°C. Mix gently before adding to centrifuge tubes. Centrifuge at 4000 x g (or higher) for 15 min. at 4°C. Use longer centrifugation time for lower speed.
  5. Collect supernatant into graduated cylinder. The supernatant should be colourless and translucent. If particulates are present, centrifugation step should be repeated and if necessary, filtered. Measure volume.
  6. Transfer supernatant into a beaker and add equal volume of cold Reagent B while stirring with stir bar. Continue stirring for 2 min. and allow suspension to stand in the cold for 1 hour.
  7. Centrifuge at 4000 x g (or higher) for 15 minutes at 4°C and discard the supernatant.  Use longer centrifugation time for lower speed.
  8. Dissolve precipitate in volume of PBS equal to original egg yolk volume and filter sterilize. IgY concentration will be between 4 and 7 mg/ml with purity of 90% or greater.
  9. Store your IgY in the refrigerator for a year (or longer) with no loss of activity.

Click here for the IgY Purification Record printout.

Technical support is available online, or call us TOLL FREE at 877- 4-GALLUS (877-442-5587)

Don't want to purify your IgY yourself?
Ask us to perform your IgY antibody purification for you.

IgY EggsPress Purification Kit

Kit Description

Our IgY EggsPress Purification Kit enables researchers to quickly and efficiently purify chicken antibodies (IgY) from the eggs of their immunized chickens without loss of activity. There are many advantages in choosing chickens to raise your polyclonal antibodies.

The disadvantage of using hens is the difficulty of purifying the IgY from the lipid environment of the egg yolk. This problem is circumvented by using our IgY antibody purification kit. Two precipitation steps — one to remove the lipid and the second to precipitate the IgY — provide you with a 90% pure IgY fraction. Expect to purify between 4 and 7 mg IgY per ml egg yolk.

Simply follow the antibody purification Kit Instructions. The entire procedure takes about 4 hours but only 30 minutes is actual hands-on time.

Once you have your IgY, you can treat it like any immunoglobulin. You may wish to label or affinity-purify your antibody. Proceed as you would with any antibody fraction. Keep in mind that IgY does not bind to protein A, protein G, or protein L! Perhaps you want to use your IgY as a primary antibody in an ELISA test, Western blot or immunostaining procedure. A selection of labeled secondary reagents (anti-chicken immunoglobulins) and normal (pre-immune) IgY are available.