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Protein Tag Purification Kit

$305.00$2,450.00

Concentration: 1 mg/ml of purified protein

Application: Recombinant Protein Purification

Reactivity: Human, Insect, Yeast

BiCell Scientific Inc has developed a series of robust protein tag purification kits allowing rapid purification of recombinant proteins from a variety of model cells.

Peptide tags can be appended to proteins so that they can be purified from their crude biological sources using affinity chromatography. Recombinant protein expression system such as E. coli, Pichia yeast, Sf9 cells and HEK293 cells, when combined with the usage of peptide tags, offers a quick and cheap solution to biomanufacturing of protein biologics.

BiCell Scientific Inc has developed a series of protein tag purification kits utilizing centrifugal columns to allow rapid purification of recombinant proteins from a variety of model cells. Each kit contains 5 pre-packed affinity columns, 5 desalting columns and a full set of buffers to complete the purification processes. These kits can achieve >90% purity without the need of sophisticated chromatography systems.

His-tag kit

  • 5 pre-packed Ni columns (bed volume: 1 ml)
  • 5 desalting columns (bed volume: 1 ml)
  • Coupling buffer (10 ml) (Tris buffered saline, pH 7.5, 10 mM imidazole)
  • Washing buffer (20 ml) (Tris buffered saline, pH 7.5, 20 mM imidazole)
  • Eluting buffer (5 ml) (Tris buffered saline, pH 7.5, 250 mM imidazole)

Store at 4oC.

GST-tag kit

  • 5 pre-packed glutathione columns (bed volume: 1 ml)
  • 5 desalting columns (bed volume: 1 ml)
  • Coupling buffer (10 ml) (Tris buffered saline, pH 7.5)
  • Washing buffer (20 ml) (Tris buffered saline, pH 7.5)
  • Eluting buffer (5 ml) (Tris buffered saline, pH 7.5, 30 mM reduced glutathione)

Store at 4oC.

Flag/Myc/HA-tag kit

  • 5 pre-packed antibody coupled columns (bed volume: 1 ml)
  • 5 desalting columns (bed volume: 1 ml)
  • Coupling buffer (10 ml) (Tris buffered saline, pH 7.5)
  • Washing buffer (20 ml) (Tris buffered saline, pH 7.5)
  • Eluting buffer (5 ml) (Glycine solution, pH 3.0)

Store at 4oC.


For Research Use Only. Not for use in clinical diagnostics.

Protein Tag Purification Procedures His-tag GST-tag Flag-tag Myc-tag HA-tag
Step 1: Equilibration

(Centrifugal condition:

1,000 xg, 5 min, room temperature)

Centrifuge pre-packed column 1x;

add 1ml of His-coupling buffer and centrifuge 2x

Centrifuge pre-packed column 1x;

add 1ml of GST-coupling buffer and centrifuge 2x

Centrifuge pre-packed column 1x;

add 1ml of  Flag-coupling buffer and centrifuge 2x

Centrifuge pre-packed column 1x;

add 1ml of  Myc-coupling buffer and centrifuge 2x

Centrifuge pre-packed column 1x;

add 1ml of  HA-coupling buffer and centrifuge 2x

Step 2: Loading samples

(Binding condition:

rotating, 30 min, room temperature)

Seal the bottom of the column;

load 1 ml of cell lysate;

cap the column and rotate the column

Seal the bottom of the column;

load 1 ml of cell lysate;

cap the column and rotate the column

Seal the bottom of the column;

load 1 ml of cell lysate;

cap the column and rotate the column

Seal the bottom of the column;

load 1 ml of cell lysate;

cap the column and rotate the column

Seal the bottom of the column;

load 1 ml of cell lysate;

cap the column and rotate the column

Step 3: Washing columns

(Centrifugal condition:

1,000 xg, 5 min, room temperature)

Centrifuge sample-loaded column 1x;

add 1ml of His-washing buffer and centrifuge 4x

Centrifuge sample-loaded column 1x;

add 1ml of  GST-washing buffer and centrifuge 4x

Centrifuge sample-loaded column 1x;

add 1ml of  Flag-washing buffer and centrifuge 4x

Centrifuge sample-loaded column 1x;

add 1ml of Myc-washing buffer and centrifuge 4x

Centrifuge sample-loaded column 1x;

add 1ml of HA-washing buffer and centrifuge 4x

Step 4: Eluting columns

(Centrifugal condition:

1,000 xg, 5 min, room temperature)

Add 0.5 ml of His-eluting buffer and centrifuge 2x;

collect elutant

Add 0.5 ml of GST-eluting buffer and centrifuge 2x;

collect elutant

Add 0.5 ml of Flag-eluting buffer and centrifuge 2x;

collect elutant

Add 0.5 ml of Myc-eluting buffer and centrifuge 2x;

collect elutant

Add 0.5 ml of HA-eluting buffer and centrifuge 2x;

collect elutant

Step 5: Buffer exchange

(Centrifugal condition:

1,000 xg, 5 min, room temperature)

Centrifuge desalting column 1x;

add 1ml of PBS and centrifuge 4x;

add 1ml of elutant and centrifuge 1x;

collect elutant

Centrifuge desalting column 1x;

add 1ml of PBS and centrifuge 4x;

add 1ml of elutant and centrifuge 1x;

collect elutant

Centrifuge desalting column 1x;

add 1ml of PBS and centrifuge 4x;

add 1ml of elutant and centrifuge 1x;

collect elutant

Centrifuge desalting column 1x;

add 1ml of PBS and centrifuge 4x;

add 1ml of elutant and centrifuge 1x;

collect elutant

Centrifuge desalting column 1x;

add 1ml of PBS and centrifuge 4x;

add 1ml of elutant and centrifuge 1x;

collect elutant

 

Additional information

Tag Purification System

His-tag, GST-tag, Flag-tag, Myc-tag, HA-tag

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Be the first to review “Protein Tag Purification Kit”

"I am really impressed with your approach. We tried multiple times previously to create monoclonal and polyclonal antibodies to claudin-2 and MLCK1. We have had limited success generating polyclonals and no success generating monoclonals. You have generated outstanding monoclonals to both. I look forward to continuing to work with you."

Jerrold R. Turner, M.D., Ph.D.

Brigham and Women’s Hospital | Harvard Medical School

"The polyclonal antibody you generated for KIAA0408 is stunning! KIAA0408 is a novel cilium molecule that has never been studied. So, clearly there will be a lot of demand for it as we have discovered a very interesting finding and the story will be published in a high impact journal. I am strongly inclined to generate monoclonal antibody for this protein too and we should think about patenting it."

Univ.-Prof. Jay Gopalakrishnan PhD

Heinrich-Heine-Universität | Universitätsklinikum Düsseldorf

"Your ARL13B antibody works beautifully!!! We’re so happy to have a cilia-specific antibody made in rat! I can send you high resolution images to be posted on your website."

Julie Craft Van De Weghe, PhD

School of Medicine | University of Washington

"The assay is a homophilic interaction mediated cell adhesion on purified protein (in this case, immobilized purified Pcdhga9 to Pcdhga9 expressed on cell surface). Compared to control, cell adhesion is reduced in the presence of Pcdhga9 monoclonal antibody supernatants!"

Divyesh Joshi, PhD

School of Medicine | Yale University

"The rabbit hybridoma supernatants of anti-APOBEC3 project are tested positive by ELISA, and we are very happy about it! We previously tried a company, Abclone. Their Project "A" has immune response that is <10,000 titer in antiserum, which would explain why there is no positive mAb after fusion. Their project "B" didn't have any immune response in rabbit."

Harshita B Gupta, PhD.

School of Medicine | UT Health San Antonio

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