Product name: Streptavidin Sepharose FF


English name: Streptavidin NUPharose Fast Flow, Streptavidin 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:


Streptavidin Sepharose FF is a bio-affinity chromatography separation medium formed by bonding Streptavidin to Sepharose microspheres. The product retains the excellent hydrophilicity of agarose and the large grid structure, and has good compatibility with biologically active macromolecules. It has the characteristics of high capacity, low non-specific adsorption, and fast flow rate.






Streptavidin Sepharose FF is based on the specific combination of streptavidin and biotin to separate and purify biotin or biotinylated antigens, antibodies, nucleic acids and other substances from biological samples or mixtures. Because streptavidin and biotin have a very strong binding force, they generally need to be separated under denaturing conditions, which can easily lead to inactivation of the sample or ligand. This feature can be used for the separation of antigen and antibody, such as binding a biotinylated antibody to a gel, and then using the affinity of the antigen-antibody to purify the non-biotinylated antigen, the antibody will not be eluted when the antigen is eluted. The reverse is also possible.






In addition, the binding of streptavidin and 2-iminobiotin is weaker than that of biotin. It is easy to bind at pH 9.5 to 11.0, and it can be eluted without denaturant at pH 4.0. Affect the activity of the sample and the activity of the gel.






Technical index:


Matrix


4% agarose gel beads


Ligand


Streptavidin


Ligand Density


~5 mg/ml


Filler particle size


60 ~ 180 μm


Maximum flow rate


800 cm/h


Recommended flow rate


15 ~ 100 cm/h


pH stability


Short-term pH 2 ~ 11; long-term pH 4 ~ 9


Back pressure resistant


0.3 MPa


Capacity


≥300 nmol/ml biotin


≥6 mg/ml biotinylated protein






Instructions:


1 Recommended buffer


1.1 Purification of biotin or biotinylated substances


Binding buffer: 20 mM NaH2PO4, 150 mM NaCl, pH 7.4


Elution buffer: 8 M guanidine hydrochloride, pH 1.5


1.2 Purification of 2-iminobiotin or 2-iminobiotin-labeled substances


Binding buffer: 50 mM ammonium carbonate, 500 mM NaCl, pH 10.0


Elution buffer: 50 mM ammonium acetate, 500 mM NaCl, pH 4.0


1.3 Purified antigen or antibody


Binding buffer: 20 mM NaH2PO4, 150 mM NaCl, pH 7.4


Elution buffer: 100 mM glycine-hydrochloric acid, pH 3.0


Note: There are two methods for the purification of non-biotinylated antigens (or biotinylated antigens to purify non-biotinylated antibodies) for biotinylated antibodies. ①The antigen can be combined with the antibody in the free state, and then when the The sample can be purified; ②The biotinylated antibody (or antigen) can also be bound to the column first, and then the corresponding non-biotinylated antigen (or antibody) can be used as the sample for purification.


2 Sample preparation


The sample on the column should be as consistent as possible with the binding buffer. The samples can usually be processed by dialysis, ultrafiltration, dilution and other methods. And before loading the column, the insoluble matter should be removed by 0.45 μm filter membrane or high-speed centrifugation.


3 Sample purification


3.1 Take an appropriate amount of Streptavidin Sepharose FF and load it into a suitable chromatography column. Equilibrate 5 column volumes with the binding buffer. The recommended flow rate is 100 cm/h.


3.2 Load the prepared sample slowly on the column. In order to ensure that the sample is fully combined with streptavidin, the sample flow rate should be controlled. The recommended flow rate is 15-50 cm/h.


3.3 After loading the sample, equilibrate more than 10 column volumes with the binding buffer, or equilibrate to the baseline to wash away impurities. The recommended flow rate is 100 cm/h.


3.4 Wash 10-20 column volumes with elution buffer. The recommended flow rate is 100 cm/h. The pH of the collected eluate should be adjusted to a stable range immediately, and the buffer should be replaced as needed.


3.5 The column after elution of the target protein should be immediately equilibrated with a neutral binding buffer, and the column should be stored in 20% ethanol, or for the next purification.

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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