IBA: Protein:Protein Interaction

Overview of Sets, Kits and Products for PPI
Gravity flow StrepMAB-Classic MacroPrep^® Column
Strep-tag II Peptide
Gravity flow StrepMAB-Immo MacroPrep^® Column

Products for PPI

For each of the described methods we offer a complete Set (for mammalian cells or E. coli) which includes all relevant products from cloning of the bait via StarGate^® (Cloning Kit) to purification of the protein complex (Purification Kit) (see table below). The kits and also single products can be purchased separately. Both a comprehensive StarGate Manual (PR26) and a PPI Manual (PR27) are delivered with the Kits and Sets..

Non-convalent prey capture

One-STrEP products
The components of the One-STrEP Set for mammalian cells and E. coli are listed in the brown column of the table below.
Please note that IBA´s former kits for mammalian cells, the "One-STrEP Starter Kit" (cat. no. 2-1109-000) and the "One-STrEP Follow Up Kit" (cat. no. 2-1110-000), are still available; these kits include the IBA Classic Vectors pEXPR-IBA103 and pEXPR-IBA105, respectively, and are thus designed for IBA´s classic cloning procedure. For cloning via StarGate^® please use the One-STrEP Sets as described here (see table below).

One-TAP products
The components of the One-TAP Sets for mammalian cells and E. coli are listed in the yellow column of the table below.
For the first purification step the same products are required as listed for the One-STrEP procedure; for the second purification step additionally a StrepMAB-Classic MacroPrep^® column and the Strep-tag^®II peptide for elution are necessary.

Two-TAP products
The components of the Two-TAP Sets for mammalian cells and E. coli are listed in the green column of the table below. For the first purification step (via the One-STrEP-tag) the same products are required as listed for the One-STrEP procedure; for the second purification step (via the FLAG-tag) an anti-FLAG M2 agarose column and a FLAG octapeptide for elution are necessary. These FLAG-products are not delivered by IBA but can be purchased from Sigma.

Convalent prey capture

SPINE products
The components of the SPINE Set for E. coli are listed in the blue column of the table below.
Cross-linking of bait and preys in vivo is achieved with p-formaldehyde which is not offered by IBA but has to be purchased elsewhere (e.g., Roth, #0335.1). For the subsequent purification of the protein complex via the Strep-tag^®II or the One-STrEP-tag the same products are required as listed for the One-STrEP procedure.

Products featuring "CMV promoter", "GST-tag", "6x Histidine-tag", "One-STrEP tag™", "StarGate^®", "Strep-Tactin^®", "Strep-tag^®", "T7 promoter", "tet promoter" are covered by patents and patent applications and Limited Use label Licenses. Please refer to http://www.iba-go.com/patents.html for the corresponding statements.
Products are for research use only.

Overview of Sets, Kits and Products for PPI

To view product description and price, and for ordering please click on cat.no.


		Mammalia	*E. coli*	Mammalia	*E. coli*	Mammalia	*E. coli*	*E. coli*
Please note that each Set contains the respective Cloning and Purification Kit.		One-STrEP Set		One-TAP Set		Two-TAP Set		SPINE Set
	cat. no.	2-1121-001	2-1121-002	2-1122-001	2-1122-002	2-1123-001	2-1123-002	2-1124-002

StarGate Kits for Cloning of Bait		One-STrEP Cloning Kit		One-TAP Cloning Kit		Two-TAP Cloning Kit		SPINE Cloning Kit
Please note that each Cloning Kit and the listed products can be purchased separately.	cat. no.	5-1621-001	5-1621-002	5-1622-001	5-1622-002	5-1623-001	5-1623-002	5-1624-002
Kit Components		Included:	Included:	Included:	Included:	Included:	Included:	Included:
StarGate Combi Cloning Set, 20 rxns, cat.no. 5-1608-000
(including competent E.coli TOP10 cells, 20 rxns)
StarGate Transfer Reagent Set, 20 rxns, cat.no. 5-1603-001
2 Acceptor Vectors, 5 rxns each; please refer to overview		pESG-IBA105	pASG-IBA105	pESG-IBA105	pASG-IBA105	pESG-IBA167	pASG-IBA167	pASG-IBA5
2 Acceptor Vectors, 5 rxns each; please refer to overview		pESG-IBA103	pASG-IBA103	pESG-IBA103	pASG-IBA103	pESG-IBA168	pASG-IBA168	pASG-IBA3
Manual PR26 (StarGate)

Kits for Purification of Protein Complex		One-STrEP Purification Kit		One-TAP Purification Kit		Two-TAP Purification Kit		SPINE Purification Kit
Please note that each Purification Kit and the listed products can be purchased separately.	cat. no.	2-1121-011	2-1121-012	2-1122-011	2-1122-012	2-1123-011	2-1123-012	2-1124-012
Gravity Flow Columns		Included:	Included:	Included:	Included:	Included:	Included:	Included:
Strep-Tactin Superflow, 5x1 ml, cat.no. 2-1207-050
Strep-Tactin Superflow, 5x0.2 ml, cat.no. 2-1207-550
StrepMAB Classic MacroPrep, 1x1 ml, cat.no. 2-1526-001
Antibody for Bait Detection
StrepMAB-Classic, HRP conjugate, 75 µl, cat.no. 2-1509-001
Buffers and Reagents
AHT, 25 mg, cat.no. 2-0401-002
Buffer L, 100 ml, 5x conc., cat.no. 2-1020-100
Avidin, 15 mg, cat.no. 2-0204-015
Buffer W, 100 ml, 10x conc., cat.no. 2-1003-100
Buffer BE, 25 ml, 10x conc., cat.no. 2-1019-025
Buffer E, 25 ml, 10x conc., cat.no. 2-1000-025
Buffer R, 100 ml, 10x conc., cat.no. 2-1002-100
Strep-tag II Peptide for elution, 5x1.8 mg, cat.no. 2-1018-009
PPI Control Donor Vector and Prey Antibody
Mammalian Control Donor Vector, 200 ng, cat.no. 2-1010-000
PP2Ac goat polyclonal antibody, 50 µl, cat.no. 2-1592-001
E.coli Control Donor Vector, 200 ng, cat.no. 2-1009-000
Manual PR27 (PPI)

Further products required for Complex Purification, but not delivered by IBA
Anti-FLAG-M2-Agarose, can be purchased from Sigma
FLAG octapeptide, can be purchased from Sigma
p-Formaldehyde, can be purchased elsewhere

Gravity flow StrepMAB-Classic MacroPrep^® Column

"One-tag tandem affinity purification" for protein:protein interaction studies

When using our One-STrEP system for protein:protein interaction analysis a second purification step is generally not required after Strep-Tactin affinity purification. For cases, in which nonetheless further purification is requested, we are offering new ready-to-use Gravity flow StrepMAB-Classic MacroPrep columns.
These columns enable a second, completely independent purification step as both the resin (MacroPrep vs Superflow) and the affinity receptor (mAB vs Strep-Tactin) are substantially different. This "one-tag tandem affinity purification" keeps the extent of bait modification minimal and thus allows better complex formation.

Specifications of Gravity flow StrepMAB-Classic MacroPrep Columns
Support	MacroPrep^® 50 (polymethacrylate)
Form	Pre-packed in 100 mM Tris/HCl pH 8.0, 1 mM EDTA, 150 mM NaCl
Strep-tag II fusion protein binding activity	> 15 nmol/ml resin
Elution	Strep-tag II fusion proteins may be gently eluted by adding 0.5 mM free Strep-tag II peptide (Cat. No. 2-1018-002) to a physiological buffer or by applying a pH 3.5 buffer.
Stability	6 months after shipment
Storage	4 °C
Shipment	RT

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Order information
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product	amount	cat. no.	order & prices
Gravity flow StrepMAB-Classic MacroPrep^® Column	1 x 1 ml	2-1526-001
Gravity flow StrepMAB-Classic MacroPrep^® Columns	5 x 0.2 ml	2-1526-505

Strep-tag II Peptide

For elution of Strep-tag proteins from StrepMAB-Classic columns

Gravity flow StrepMAB-Immo MacroPrep^® Column

Columns used e.g. for sequential elution of prey proteins in protein:protein interaction studies

StrepMAB-Immo, a murine monoclonal antibody, binds SerAla-One-STrEP-tag (and SerAla- Strep-tag II) fusion proteins nearly irreversibly. Thus, these columns are designed for the stable and oriented immobilization of bait proteins. Analysis of interacting proteins is performed by sequential elution of prey proteins from the bait via Gravity flow StrepMAB-Immo MacroPrep Columns using adequate buffers. The bait proteins are presumed to stay bound under a wide range of buffer conditions.
Please note, that Gravity flow StrepMAB-Immo MacroPrep Columns are not recommended for the purification of One-STrEP-tag or Strep-tag II fusion proteins because elution is not possible under physiological conditions.

*Specifications of Gravity flow StrepMAB-Immo MacroPrep Columns*
Specifications of Gravity flow StrepMAB-Immo MacroPrep Columns	Nearly irreversible binding of C-terminally fused Strep-tag II with SerAla linker (rec. protein seq.- SerAla-TrpSerHisProGlnPheGluLys). The Strep-tag II will be linked automatically via the SerAla dipeptide to the C-terminus when the vectors for C-terminal fusion from IBA are used. Other linker sequences than SerAla lead to reduced affinity in the cases tested so far. The sequence SAWSHPQFEK fused to the N-terminal end of a recombinant protein has not been tested so far and, therefore, it is uncertain whether this equally mediates nearly irreversible binding.
Support	MacroPrep^® 50 (polymethacrylate)
Form	Pre-packed in 100 mM Tris/HCl pH 8.0, 1 mM EDTA, 150 mM NaCl
Strep-tag II fusion protein binding activity	> 15 nmol/ml resin
Stability	6 months after shipment
Storage	4 °C
Shipment	RT