Alkali-tolerant Recombinant Protein A Agarose

Catalog Number:
LT12014
Packing Details:
1 mL resin, crosslinked 6% beaded agarose supplied as 50% slurry (e.g., 1mL of settled resin is equivalent to 2mL of 50% slurry) in 0.02% sodium azide
Binding Capacity:
10~20 mg/ml rabbit IgG or 15~25 mg/ml human IgG
Support pH Stability:
2-11 (short term); 3-10 (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: 30cm 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 A Agarose is an alkali-tolerant Protein A Agarose that has been covalently immobilized onto 6% beaded agarose. The high binding Protein A agarose resin can be used for antibody affinity purificaiton. The immobilized Protein A is used for polyclonal IgG purification from rabbit, human, dog or cat serum.

Highlights:

  • Alkali tolerant: Improved alkaline resistance. NaOH can be used to regenerate resin.
  • High Binding Capacity: The high binding capacity of 10-20 mg/ml IgG to Protein A is perfect for batch- or column- type affinity purification.
  • Stability: Specially designed sepharose linked to recombinant alkali-tolerant Protein A with spacer arms for improved stability and binding capability.

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

Elution buffer: 100 mM sodium citrate, pH 3.0

Neutralization buffer: 1M citrate buffer, pH 9.0

  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 A agarose gel.

Recombinant Staphylococcal Protein A Agarose          Recombinant Staphylococcal Protein A Agarose

Typical binding and elution conditions with protein A Agarose

Species

Antibody Class

Protein A 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

++

 

2

Goat

IgG2

+

 

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

IgG3

+

8.0~9.0

3.0~4.0

Rabbit

no distinction

++++

7.4

3.5~4.5

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!
  • 1 Units in Stock

$280.00

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