Welcome
Catalogue 2012
About us
Contact us
How to Order
Proteomic Research
IMAC Purification
How to remove 'heavy metal ions' from your protein solution after IMAC elution steps?
PDC
PDC Ordering
IgA, IgG, IgM, IgY Purification Kits
IgY Purification Kit
IgG, IgM, IgY Immobilization Kits
Concentration Resin
Affinity media
Affinity Purified Proteins & Hormones
Antibodies
Sample Clean Up and Immunoaffinity Capture
Customer Services
Links
Distribution
PDF Catalogue
ISO 9001:2008
Scientific contributions
News
Affiland

  PDF version

Polyclonal Sheep IgG purification kit

Product code: SIKPG-FF KIT

under physiological conditions, all steps

PRICE: 300 EUR/KIT

KIT CONTENT
(sufficient for 12 purifications of IgG from 15 ml of Sheep serum)

  • Sheep IgG Binding Gel (SepharoseTM fast flow) (Code : SIKPG-FF) : 5 ml gel column.
    Binding capacity : > 20 mg Sheep IgG/ml wet gel.
    Purity : 98% by SDS-PAGE.
    Maximum pressure : 3 bars (43 psi, 0.3 MPa).
  • Sheep IgG Loading Buffer (Code : LBSPG) : 1.5x concentrated : 1,000 ml. Add 500 ml of distilled water to have a total of 1500 ml before use.
  • Sheep IgG Binding Buffer (Code : BBSPG) : The Sheep IgG Binding Buffer (Code : BBSPG) will be obtained by adding 1 volume of distilled water to 1 volume of Sheep IgG Loading Buffer (Code : LBSPG).
  • Sheep IgG Elution Buffer (Code : EBSPG) : 8x concentrated : 125 ml. Add 875 ml of distilled water to have a total of 1000 ml before use.

INSTRUCTION FOR USE

  1. Equilibrate the column with 15 ml of Sheep IgG Binding Buffer (BBSPG).
  2. Prepare the sample by dissolving 15 ml of Sheep serum in 15 ml of Sheep IgG Loading Buffer (Code : LBSPG).
  3. Load the sample prepared in point 2 into the column prepared in point 1 at a flow rate of 30 ml/hour.
  4. Wash the column with 180 ml of Sheep IgG Binding Buffer (BBSPG).
  5. Elute the Sheep IgG with 50 ml of Sheep IgG Elution Buffer (Code : EBSPG) until the O.D. at 280nm of the eluent reaches the baseline level.
  6. Assay the elution fractions obtained as described in point 5, using the most appropriate system (SDS-PAGE, immunodiffusion, radioimmunoassay, Elisa...)
REGENERATION OF THE SHEEP IgG BINDING GEL
It is recommended to regenerate the gel after every 5 cycles of use.
  1. Wash the column with 10x volumes of NaOH 0.1M.
  2. Wash the column with 10x volumes of distilled water.
  3. Equilibrate the column 10x volumes of PBS (50 mM K2HPO4, 150mM NaCl) pH 7.4.
  4. Store the column at 4°C in the presence of NaN3 0.1% (w/v).
  5. For the next use, see INSTRUCTION FOR USE as described above.
If you need sterile materials, the regeneration can be carried out as follows.
STERILE REGENERATION OF THE COW IgG BINDING GEL
(GEL SANITIZATION)
AFTER EVERY 5 CYCLES OF USE
  1. Wash 1 volume of gel column with 5 volumes of acetic acid 1 M.
  2. Wash this column with 10 volumes of sterile distilled water.
  3. Wash this column with 5 volumes of NaOH 1M.
  4. Wash this column with 10 volumes of sterile distilled water.
  5. Wash this column with 10 volumes of PBS (50 mM K2HPO4, 150mM NaCl) pH 7.4; NaN3 0.1%(w/v).
  6. The sterile gel column is now ready to be re-used.

  IgA, IgG, IgM, IgY Purification Kits