Product Name: Recombinant Protein A Sepharose FF


English name: Recombinant Protein A NUPharose Fast Flow, rProtein A NUPharose FF


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


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


Storage: 20% ethanol, 2 ~ 8℃


Shelf life: 3 years


Related introduction:


Protein A is a Staphylococcus aureus cell wall protein that contains 5 regions that can bind to the Fc fragment of IgG. As an affinity ligand, protein A can be more tightly coupled to NUPharose, so that these regions can freely bind to free IgG. One molecule of protein A can bind two molecules of IgG.






Technical index:


Matrix


4% highly cross-linked agarose gel


Ligand


Recombinant Protein A


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


Back pressure resistant


0.3 MPa


Capacity


≥40 mg/ml IgG






The binding power of antibody to protein A and protein G:


type


Species


Subtype


Antibody Class


Protein A


Protein A


Protein G


Protein G


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.






Application examples:


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


Elution buffer: 0.1 M citrate buffer, pH 4.0


Neutralization buffer: 1 M Tris ~ HCl, pH 9.0


1) 1 ml of recombinant protein A sepharose 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 5 ml of rabbit polyantiserum to 50 ml with binding buffer, filter and load the sample with a 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 each use, regenerate and clean with 3 ~ 5 column volumes of 0.1 M citrate buffer (pH 3.0);


7) The purity of the rabbit polyclonal antibody eluted by SDS-PAGE (Figure 1) electrophoresis analysis is above 95%.






Picture 44.jpg


figure 1






Precautions:


1) The pre-packed column is easy to use and can complete the purification task 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 in the collection container 5% ~ 20%, pH 7 ~ 9 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 and affect the purification effect. If bubbles have already occurred, you can repack the column by yourself.


4) Normally (every time) after the target sample is eluted, continue to wash 3 ~ 5 column volumes with 0.1 M citrate buffer (pH 2 ~ 3), and then wash more than 5 column volumes with equilibration buffer for simple regeneration .


5) In-place cleaning should be performed after several times of use. It is recommended to wash 1 to 3 column volumes (10 to 30 min) with 0.1% Triton X-100 at 37°C, and immediately wash more than 5 column volumes with equilibration buffer; or 70 Wash with% ethanol for more than 5 column volumes (can be maintained for 4 to 16 h), and then wash with equilibration buffer for more than 5 column volumes. Cleaning-in-place can remove strong hydrophobic proteins and lipids and restore the load and flow rate of the column.


6)


Recommended flow rate


Pre-packed column volume (ml)


1 ml


5 ml


10 ml


Flow rate (ml/min)


0.2


1

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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.