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Product Name: Recombinant Protein L Sepharose FF


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


Specifications: 20 ml, 100 ml, 1L, 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:


Recombinant Peptostreptococcus magnus Protein L (r-PPL) is coupled to a highly cross-linked agarose gel with an affinity purification medium. Peptococcus magensis protein L (PPL) can bind to the κ light chain of the antibody without affecting the antigen binding site of the antibody. This feature makes it more extensive than the protein A and protein G that bind to the Fc region of the antibody. Combine antibodies from various sources and subclasses. Such as human IgG, IgM, IgA, IgE, IgD, and Fab, scFv and other antibody fragments containing kappa light chain (human type 1, 3, 4, mouse type 1). But it cannot bind to heavy chain, lambda light chain, kappa light chain (type 2), so recombinant protein L Sepharose FF cannot purify antibodies from cows, goats, sheep, and chickens.






Technical index:


Matrix


4% highly cross-linked agarose gel


Ligand


Recombinant protein L


Ligand Density


≥6 mg/ml


Filler particle size


60~180 μm


Maximum flow rate


800 cm/h


Recommended flow rate


20-100 cm/h


Working (cleaning) pH


2~9(2~10)


Back pressure resistant


0.3 MPa


Capacity


≥40 mg/ml human IgG






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


type


Species


Subtype


Antibody Class


Protein A


Protein A


Protein G


Protein G


Protein L


Protein L


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)


+


+


-


-


-


+++


NA


NA


NA


Rabbit


IgG


+++


+++


+


Pig


IgG


+++


+


+++


Dog


IgG


+++


+


NA


Chicken


IgY


-


-


-


Note:-means no binding; + means weak binding; ++ means medium binding; +++ means very strong binding; NA means no data yet. Antibodies that can be used for affinity purification generally require moderate or higher binding power.






Application examples:


Binding buffer: 20 mM phosphate buffer (PB), pH 7.4


Elution buffer: 0.1 M citric acid-sodium citrate buffer, pH 2.7


Neutralization buffer: 1 M Tris-HCl, pH 9.0


1) 1 ml Recombinant Protein L Sepharose FF pre-packed column, wash away the preservation solution with 5-10 column volumes of distilled water;


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


3) Dilute 3 ml of mouse ascites with binding buffer to 15 ml, filter and load with 0.45 μm filter membrane;


4) Equilibrate 5-10 bed volumes with binding buffer to the baseline;


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


6) Regenerate and clean with 3~5 column bed volumes of elution buffer after each use;


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










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, which will affect the purification effect. If bubbles have already occurred, you can repack the column by yourself.


4) Normally, after the target sample is eluted, continue to wash 3~5 column volumes with 0.1 M citric acid-sodium citrate buffer (pH 2.0~2.7), and then wash more than 5 column volumes with equilibration buffer for simple regeneration .


5) Wash in place after several times of use. It is recommended to wash 1-3 column volumes (10~30min) with 0.1% Triton X-100 at 37℃, and immediately wash more than 5 column volumes with equilibration buffer; or 70% Wash with ethanol for more than 5 column volumes (can be kept for 4-16h), then wash with equilibration buffer for more than 5 column volumes; or wash with 10mM NaOH for 1-3 column volumes (10-20min), and then wash 5 columns with equilibration buffer Above the volume. Cleaning-in-place can remove strong hydrophobic proteins and lipids and restore the load and flow rate of the column.

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.