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

Ask a Question

$420.00

Add to Cart:

Customers who bought this product also purchased...

LifeTein LifeTein provides custom peptide synthesis service, recombinant proteins, peptides, cytokines, custom antibody service and custom protein service. LifeTein is the world leader in fast peptide synthesis service with lab facility located in New Jersey USA. LifeTein Peptide 100 Randolph Road, Suite 2D, Somerset USA New Jersey 08873
Copyright © 2024 LifeTein.com. Powered by LifeTein