Recombinant Protein G Agarose

Catalog Number:
LT12015
Packing Details:
1 mL resin, crosslinked 6% beaded agarose supplied as 50% slurry (e.g., 2mL of settled resin is equivalent to 4mL of 50% slurry) in 0.02% sodium azide, ligand density: 4-5mg/ml
Binding Capacity:
≥50 mg/ml rabbit IgG or ≥50 mg/ml human IgG
Support pH Stability:
2-14 (short term); 3-13 (long term)
Average Particle Size:
45 to 165 microns
Exclusion Limit:
10,000 to 4,000,000 daltons
Maximum Volumetric Flow Rate:
approx. 1mL/minute (for 1cm diameter column); Maximum Linear Velocity: 300 cm per hour; Maximum Pressure: less than 4.5psi (0.3 bar)
Storage:
4 °C - 8 °C in 20% ethanol
Shelf Life:
3 years
Description:

LifeTein Protein G Agarose consists of recombinant Protein G that has been covalently immobilized at high density onto high-quality crosslinked 6% beaded agarose. Protein G is especially suited for use with mouse antibodies in addition to most IgG isotypes from human, goat, cow and sheep sera, including human IgG3 and mouse IgG1 isotypes.

Protein G is a bacterial cell wall protein original from group G Streptococcus and now produced as a recombinant in E. coli. Recombinant Protein G has a mass of approximately 31 to 34 kDa by SDS-PAGE. IgG-binding function is optimal at pH 5 but also occurs efficiently in near-neutral conditions (pH 7.0 to 7.2). Protein G binds to most mammalian immunoglobulins primarily through their Fc regions.

Compared to Protein A, Protein G binds a broader spectrum of IgG subclasses from human, mouse and rat serum. Protein G exhibits stronger binding to human IgG3, mouse IgG1 and all three isotypes of Rat IgG. However Protein G binds more weakly than Protein A to pig, guinea pig, dog and cat IgG.

Protocol:

Protein G Binding buffer: 20 mM sodium phosphate, 150 mM NaCl, pH 7.4

Protein G Elution buffer: 100 mM Glycine, pH 3.0

Protein G Neutralization buffer: 1M Tris buffer, pH 7.5-9

  1. Wash the prepacked column with 5~10 column volumes of distilled water to remove 20% ethanol.
  2. Equilibrate the column with 5~10 column volumes of binding buffer.
  3. 1:10 dilution of serum with binding buffer. Filtrate the diluted serum through a 0.45 μm filter and load the sample.
  4. Wash with 10 column volumes of binding buffer.
  5. Elute with 5 column volumes of elution buffer and neutralize collect fractions with neutralization buffer.
  6. After each separation cycle, regenerate the resin by washing with approximately 3~5 column volumes of 0.1 M citrate buffer (pH 3.0).
  7. Confirm the purity of the collected antibody by SDS-PAGE analysis.
  8. Purification capacity: ≥30 mg of rabbit IgG per ml of recombinant protein G agarose gel.

Recombinant Staphylococcal Protein A Agarose          Recombinant Staphylococcal Protein A Agarose

Typical binding and elution conditions with protein G Agarose

Species

Antibody Class

Protein G binding

pH value of binding buffer

pH value of elution buffer

Human

IgG1

+++

6.0~7.0

3.5~4.5

IgG2

+++

6.0~7.0

3.5~4.5

IgG3

+++

8.0~9.0

<7.0

IgG4

+++

7.0~8.0

2.5~4.5

Cow

IgG2

+++

7.0~8.0

2

Goat

IgG2

+++

7.0~8.0

5.8

Mouse

IgG1

++

8.0~9.0

5.5~7.5

IgG2a

+++

7.0~8.0

4.5~5.5

IgG2b

+++

7.0

3.5~4.5

IgG3

+++

7.0

4.0~7.0

Rat

IgG1

+

>9.0

7.0~8.0

IgG2a

+++

>9.0

<8.0

IgG2b

+

>9.0

<8.0

IgG2c

++

8.0~9.0

3.0~4.0

Rabbit

IgG

+++

7.4

2.7~4.0

Notes:
  1. Avoid air bubbles.
  2. Regenerate the resin every 5 purification in order to maintain the product efficiency.
  3. After every 10 separation cycle, wash the resin with 5 column volumes of 20% ethanol and 5~10 column volumes of 70% ethanol sequentially to remove hydrophobic substances.
  4. Keep at 4~8 °C in 20% ethanol.
For research use only!
  • 8 Units in Stock

$350.00

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