Recombinant Protein G Sepharose FF Gravity Prepacked Column
| 【Numbering】BK-NRPB02S | 【CAS】CAS | ||
| 【Item No.】BK-NRPB02S-Z25 | 【specification】 | ||
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Product Name: Recombinant Protein G Sepharose FF
English name: Recombinant Protein G NUPharose Fast Flow, rProtein G NUPharose FF
Catalog number: NRPB02L, NRPB02S (prepacked column)
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:
Streptococcus Protein G (SPG) is a protein in the cell wall of G and C streptococci. It can specifically bind to the Fc segment of immunoglobulin IgG molecules without affecting the binding of Fab segments to antigen molecules. Ability, its affinity filler can be used to bind, adsorb and purify immunoglobulins (antibodies) from serum, ascites, tissue culture and other supernatants.
Technical index:
|
基質 |
4%高度交聯瓊脂糖凝膠 |
|
配基 |
重組蛋白G |
|
配基密度 |
≥6 mg/ml |
|
填料粒徑 |
60 ~ 180 μm |
|
最大流速 |
800 cm/h |
|
推薦流速 |
20 ~ 100 cm/h |
|
工作（清洗）pH值 |
2 ~ 9(2 ~ 10) |
|
耐反壓 |
0.3 MPa |
|
載量 |
≥40 mg/ml IgG |
抗體與蛋白A、蛋白G的結合力：
|
種類 Species |
亞型 Antibody Class |
蛋白A Protein A |
蛋白G Protein G |
|
人 Human |
IgG1 IgG2 IgG3 IgG4 IgM IgD IgA Fab |
+++ +++ + +++ + - + + |
+++ +++ +++ +++ - - - + |
|
小鼠 Mouse |
IgG1 IgG2a IgG2b IgG3 IgM |
+ +++ +++ +++ - |
++ +++ +++ +++ - |
|
大鼠 Rat |
IgG1 IgG2a IgG2b IgG2c |
+ - - +++ |
++ +++ + +++ |
|
牛 Cow |
IgG1 IgG2 |
+ +++ |
+++ +++ |
|
山羊 Goat |
IgG1 IgG2 |
+ +++ |
+++ +++ |
|
馬 Horse |
IgG(ab) IgG(c) IgG(T) |
+ + - |
- - +++ |
|
兔Rabbit |
IgG |
+++ |
+++ |
|
豬Pig |
IgG |
+++ |
+ |
|
狗Dog |
IgG |
+++ |
+ |
|
雞Chicken |
IgY |
- |
- |
Note:-means no combination; + means weak combination; ++ means medium combination; +++ means very strong combination.
Application examples:
Binding buffer: 20 mM phosphate buffer (PB), pH 7.4
Elution buffer: 0.1 M glycine-hydrochloric acid buffer, pH 2.7
Neutralization buffer: 1 M Tris-HCl, pH 9.0
1) 1 ml recombinant protein G sepharose gel pre-packed column, wash away the preservation solution with 5-10 column volumes of distilled water;
2) Equilibrate 5-10 bed volumes with binding buffer;
3) Dilute 5 ml of rabbit polyantiserum to 50 ml with binding buffer, filter and load with 0.45 μm filter membrane;
4) Equilibrate 5-10 column bed volumes with binding buffer to 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 rabbit polyclonal antibody eluted by SDS-PAGE (Figure 1) electrophoresis analysis is above 95%.
圖 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, after the target sample is eluted, continue to wash 3 to 5 column volumes with 0.1 M glycine-hydrochloric acid buffer (pH 2.0 ~ 2.7), 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 4-6 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.
| 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. |
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