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Rat IgG purification kit

Product code: RIK-FF KIT

PRICE: 300 EUR/KIT

KIT CONTENT
(sufficient for 8 purifications with 15 ml rat serum/each)

  • Rat IgG Binding Gel (SepharoseTM fast flow) (Code : RIK-FF) : 5 ml gel column.
    Binding capacity : approx. 12 mg Rat IgG/ml wet gel.
    Purity : 95% by SDS-PAGE, free of albumin.
    Maximum pressure : 3 bars (43 psi, 0.3 MPa).
    Gel life : approx. 50 cycles with routine regeneration.
  • Rat IgG Binding Buffer (Code : BBR) : 2 x 1,050 ml.
  • Rat IgG Elution Buffer (Code : EBR) : 500 ml.
  • Rat IgG Precipitating Agent (Code : PAR) : 8 x 1 sachet of sufficient quantity for precipitating all IgG from 15 ml of Rat serum.
 

INSTRUCTION FOR USE

  1. Add with mild agitation 1 sachet of Precitating Agent (PAR) 15 ml of Rat serum for 10 minutes. Stop the agitation and allows to stand for 30 minutes at 4°C. Centrifuge at 3000 g for 10 minutes. Discard the supernant from the pellet. Dissolve the pellet in 30 ml of Rat IgG Binding Buffer (BBR). Such a sample is ready to be loaded into the column.
  2. Equilibrate the column with 20 ml of Rat IgG Binding Buffer (BBR). Set the valve to get a flow rate of approx. 30 ml/hour.
  3. Load the sample prepared in point 1 into the column prepared in point 2 at a flow rate of 30 ml/hour.
  4. Wash the column with 200 ml of Rat IgG Binding Buffer (BBR) at a flow rate of approx. 50 ml/hour.
  5. Elute the Rat IgG with the Rat IgG Elution Buffer (Code : EBR) until the O.D. at 280nm of the eluent reaches the baseline level. Collect 10 fractions of 5 ml elution volume.
  6. Assay the elution fractions obtained as described in point 5, using the most appropriate system (SDS-PAGE, immunodiffusion, radioimmunoassay, Elisa...)
REGENERATION OF THE RAT 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 RAT IgG BINDING GEL
(GEL SANITIZATION)
AFTER EVERY 5 CYCLES OF USE
  1. Wash the column with 5 volumes of acetic acid 1 M.
  2. Wash the column with 10 volumes of sterile distilled water.
  3. Wash the column with 5 volumes of NaOH 1M.
  4. Wash the column with 10 volumes of sterile distilled water.
  5. Wash the 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.
Rat

  IgG, IgM, IgY Purification Kits