Recombinant Protein A Sepharose FF
| 【Numbering】BK-NRPB01L | 【CAS】CAS | ||
| 【Item No.】BK-NRPB01L-100 | 【specification】 | ||
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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, so you should immediately neutralize the collected antibody solution to neutral with an alkaline neutralization buffer (such as 1 M Tris-HCl, pH 9.0), or put it in a collection container beforehand. 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) Usually (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) Wash in place 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
2
Message
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, so you should immediately neutralize the collected antibody solution to neutral with an alkaline neutralization buffer (such as 1 M Tris-HCl, pH 9.0), or put it in a collection container beforehand. 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) Usually (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) Wash in place 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
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 |
Message
|
| 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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