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Product name: Protein A/G Sepharose FF


English name: Protein A/G NUPharose Fast Flow, Protein AG NUPharose FF


Catalog number: NRPB13L, NRPB13S (prepacked column)


Specifications: 20 ml, 100 ml, 1L, 1 ml pre-packed column, 5 ml pre-packed column, special specifications are customized


Transportation: 2 ~ 25℃, normal pressure, avoid light


Storage: 20% ethanol, 2 ~ 8℃


Shelf life: 3 years






Related introduction:


Protein A Protein G Sepharose FF is an affinity separation medium formed by bonding recombinant Protein A Protein G fusion protein (r-PAG, NRPA14S) to Sepharose microspheres. Compared with separate protein A or protein G separation medium, it has a wider binding range and can bind to all human IgG subtypes, IgA, IgE, IgM, but cannot bind to mouse IgA, IgM and serum albumin , Suitable for the extraction and detection of mouse IgG monoclonal antibodies. At the same time, the fusion of r-PAG reduces the dependence on pH, allowing binding at pH 5 ~ 8.






Technical index:


Matrix


4% cross-linked agarose gel microspheres


Ligand


Protein A protein G fusion protein


Ligand Density


≥6 mg/ml


Filler particle size


60 ~ 180 μm


Maximum flow rate


800 cm/h


Recommended flow rate


20 ~ 100 cm/h


pH stability


pH 3 ~ 9, pH2 ~ 10 (cleaning)


Back pressure resistant


0.3 MPa


Capacity


≥ 20 mg/ml IgG


Storage Conditions


Stored in 20% ethanol solution at 2 ~ 8℃






Application examples:


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


Elution buffer: 0.1 M Glycine-HCl buffer, pH 2.5


Neutralization buffer: 1 M Tris-HCl, pH 9.0


1) 1 ml Protein A Protein G Sepharose FF pre-packed column is washed with 5-10 bed volumes of distilled water to remove the preservation solution;


2) Equilibrate 5-10 bed volumes with binding buffer and buffer;


3) Dilute 3 ml of rabbit polyantiserum to 15 ml with binding buffer, filter the sample with 0.45 μm filter;


4) Wash with binding buffer for another 5-10 bed volumes;


5) Elute the antibody with elution buffer, collect the elution peak, and adjust its pH to neutral with neutralization buffer;


6) After the target antibody is eluted, wash 5-10 column volume equilibration columns with binding buffer;


7) Analyze the eluted rabbit polyclonal antibody.


type


Species


Subtype


Antibody Class


Protein A


Protein A


Protein G


Protein G


Protein A protein G fusion protein


Protein AG


 


 


 


people


Human


IgG1


IgG2


IgG3


IgG4


IgM


IgD


IgA


Fab


+++


+++


+


+++


+


-


+


+


+++


+++


+++


+++


-


-


-


+


+++


+++


+++


+++


+


-


+


+


 


Mouse


Mouse


IgG1


IgG2a


IgG2b


IgG3


IgM


+


+++


+++


+++


-


++


+++


+++


+++


-


++


+++


+++


+++


-


 


Rat


Rat


IgG1


IgG2a


IgG2b


IgG2c


+


-


-


+++


++


+++


+


+++


++


+++


+


+++


Cattle


Cow


IgG1


IgG2


+


+++


+++


+++


+++


+++


goat


Goat


IgG1


IgG2


+


+++


+++


+++


+++


+++


Horse


Horse


IgG(ab)


IgG(c)


IgG(T)


+


+


-


-


-


+++


+


+


+++


Rabbit


IgG


+++


+++


+++


Pig


IgG


+++


+


+++


Dog


IgG


+++


+


+++


Chicken


IgY


-


-


-


Note:-means no combination; + means weak combination; ++ means medium combination; +++ means very strong combination.






Precautions:


1) The pre-packed column is simple and convenient to use, and the purification task can be completed without equipment;


2) After the sample is eluted, the pH is generally very low, and the collected antibody solution should be neutralized to neutral with an alkaline neutralization buffer (such as 1 M Tris-HCl, pH 9.0) immediately, or pre-filled 5% ~ 10%, pH 7.5 buffer (such as 1 M Tris-HCl or 1 M phosphate buffer) to help maintain the biological activity of the antibody and avoid antibody inactivation;


3) When using, ensure that the temperature of the column and the buffer are the same to avoid bubbles in the column bed. If bubbles have already occurred, you can repack the column by yourself;


4) After repeated use, there will be precipitated proteins, strong hydrophobic proteins, lipoproteins, liposomes and other non-specific adsorption gels, which can be cleaned with 0.1% detergent for a short time, such as 0.1% Triton X-100 Column volume, immediately wash 5-10 column volumes with binding buffer. It can also be cleaned with 70% ethanol for the same cleaning.


5)


Recommended flow rate


Pre-packed column volume (ml)


1 ml


5 ml


10 ml


Flow rate (ml /min)


0.2


1


2 

Warm tips: Suzhou Beike nano products are only used for scientific research, not for human body,different batches of products have different specifications and performance

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Warm tips: Suzhou Beike nano products are only used for scientific research, not for human body,different batches of products have different specifications and performance.The website pictures are from the Internet. The pictures are for reference only. Please take the real object as the standard. In case of infringement, please contact us to delete them immediately.