antibody Protein A Agarose Resin for polyclonal antibodies purification

Delivery Time:Depand on stock, without stock need 4 weeks for produce.
Certification:ISO9001:2008

Recombinant Protein A Sepharose Resin for polyclonal antibodies purification ,antibody , Protein A Agarose

Product Name: Alkali-tolerant Protein A agarose FF
Catalog Number:NRPB06L, NRPB06S (prepacked column)
Packing details: 10 ml, 100 ml, 1L, 1 ml(prepacked column), 2 ml(prepacked column), 5ml(prepacked column), 10ml(prepacked column)
Storage condition: 4oC

Company Group:                                                                                                                      Hangzhou NeuroPeptide Biological Science and Technology Incorporation, Ltd. (NUPTEC----Sales Department)

Add:   Room 11-08, HuaRui Building, No.66, Jianshe Road 1st, Xiaoshan,

          Hangzhou, Zhejiang, China 311202

Hangzhou NUPTEC Rising Bioproducts Incorporation, Ltd. ( NUPTEC----GMP Manufactory)

Add:   600 21st Boulevard, Xiasha Economy and Technology District,

          Hangzhou, China 310018

Introduction:

Protein A is a cell wall protein of Staphylococcus aureus and contains five functional domains that can bind to the Fc fragment of IgG. Structural modification of nature protein A improved its alkaline resistance. Therefore, NaOH could be used for removal of pyrogen and cleaning. The advantages of a spacer arm of agarose combined with the directed binding sites of recombinant protein A ensure the oriented coupling between agarose and recombinant protein A as well as the high affinity of recombinant protein A with substances.

Column characteristics:

Support	6% highly cross-linked spherical agarose
Ligand	rat-PA
Ligand Density	5~6 mg/ml
Particle Size	45~165 μm
Maximum Flow Rate	700 cm/h
Recommended Linear Flow Rate	20~100 cm/h
pH Limits	pH 3~13
Maximum Operating pressure	0.3 MPa
Capacity	≥50 mg/ml rabbit IgG or ≥50 mg/ml human IgG
Storage	4 oC to 8 oC in 20% ethanol

Performing a separation:

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

Elution buffer: 100 mM sodium citrate, pH 4.0

Neutralization buffer: 1M Tris-HCl, pH 9.0

1) Wash the prepacked 1 ml 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) Dilute 5 ml rabbit serum with binding buffer to 50 ml. Filtrate the diluted serum through a 0.45 レm 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 NaOH(pH 3.0).

7)Confirm the purity of the collected antibody by SDS-PAGE analysis (Figure 1, >95% purity).

Figure 1

M- Protein Molecular Weight Marker

1-Rabbit Antibody (non-reduced)

2-Rabbit antibody (reduced)

Notice:

1)     It is easy and simple to use the prepacked column. It is not necessary to need any equipment to purify antibody.

2)     Be sure that the antibody is stable under the elution conditions selected. If there is any doubt, titrate the antibody fraction to neutrality immediately upon elution in order not to lose biological activity. To accomplish this, prefill the fraction vessels to 5~10% of the intended fraction volume with a buffer concentrate at about pH7.5 (for example 1 M Tris-HCl or 1 M sodium phosphate).

3)     Make sure temperature of the equilibration buffer identical to that of the packed column to avoid air bubbles. If air has entered the column, the column should be repacked.

4)     It is recommended to regenerate the resin once after 5 times separation cycle in order to extend the service life.

5)     After 10 times separation cycle, wash the resin with 5 column volumes of 0.1~0.5M NaOH and 5~10 column volumes of 70% ethanol sequentially to remove hydrophobic substances.

6)     For longer periods of storage, keep at 4~8oC in 20% ethanol.

NUPTEC EXHIBITION History: 

Time:December 13-15, 2012           1st Shenzhen International Biotech Innovation Forum & Exhibition

Time: March 13-15, 2013                 PCHI2013 Personal Care and Homecare Ingredients

Time: March 18-20, 2013                 BIT’s 5th Annual International Congr