Antibodies & Strep-Tactin(Antibodies)

Description
Application & Reference
Specifications

Strep-tag® antibodies

The following monoclonal antibodies against Strep-tag® are available:

StrepMAB-Classic, unconjugated
StrepMAB-Classic HRP conjugate (HRP, horseradish peroxidase)
StrepMAB-Immo - the high affinity antibody for capturing Strep-tag® proteins on solid surfaces

Further products:

Strep-tag® Protein Ladder as positive control for Western blots and Molecular Weight Marker
Fluorescent Detection with our Trial Kits for different wavelength

StrepMAB-Classic

StrepMAB-Classic is recommended for the following applications:

Immunofluorescence
FACS
Detection in ELISA

StrepMAB-Classic, horseradish peroxidase (HRP) conjugate

The StrepMAB-Classic, HRP conjugate¹ is recommended for detection of Strep-tag® II proteins in Western blots without the need for secondary antibodies.

¹The conjugate is useful for both chemiluminescence and chromogenic detection.

***Strep*MAB-Immo**

High-affinity Strep-tag® II specific monoclonal antibody

high affinity both with C- and N-terminal Strep-tag® fusion, provided the Strep-tag® is N-terminally extended by a SerAla-linker
for FACS analysis and Immunofluorescence staining
optimal for capturing Strep-tag® fusion proteins on solid phases(e.g. Biacore chips¹)
StrepMAB-Immo coated microplates for immobilization of Strep-tag® proteins

For Western blots use StrepMAB-Classic HRP conjugate.

StrepMAB-Classic/-Immo fluorescent conjugates

For fluorescent detection of Strep-tag^® proteins in

Immunocytochemistry and immunohistochemistry
FACS analysis

Available as single conjugates of 50µg for different wavelength:

StrepMAB Classic or StrepMAB Immo conjugated with Chromeo 488™

StrepMAB Classic or StrepMAB Immo conjugated with Chromeo 542

StrepMAB Classic or StrepMAB Immo conjugated with Oyster 645

Note, Fluorescent Detection Trial Kits were discontinued on December 31, 2013 and replaced by single fluorescent conjugates.

Secondary HRP antibody

Rabbit anti Mouse pAb, HRP conjugated

Rabbit anti Mouse pAb, horseradish peroxidase (HRP) conjugated

Rabbit anti Mouse pAb, HRP conjugated, is recommended for detection of

bound mouse monoclonal antibodies, like StrepMAB-Classic, in Western blots (chromogenic reaction),
mouse monoclonal antibodies, like StrepMAB-Classic, in microplate assays

Strep-tag® Protein Ladder

Accurate MW determination and positive control on Western blots!

Strep-tag® Protein Ladder

Accurate MW determination and positive control on Western blots!

The Strep-tag® Protein Ladder(#2-1011-100) simplifies the comparison of Coomassie stained gels with corresponding Western blots. Thus, Strep-tag® fusion proteins can be located accurately in complex band patterns on Coomassie gels.

The Strep-tag® Protein Ladder is a mixture of six recombinant, highly purified Strep-tag® proteins employed for precise sizing of proteins by SDS-PAGE.

The proteins resolve into clearly identifiable sharp and evenly stained bands from 15 to 100 kDa when analyzed on an SDS gel and stained with Coomassie Blue.

As each protein contains the Strep-tag® II sequence which is detected by Strep-Tactin® conjugates or Strep-tag® specific antibodies, the ladder can also be used for MW determinations on Western blots and serves as a positive control for the various detection systems.

StrepMAB-Classic

Immunofluorescence

Strep-tag-FasL fusion protein stained with StrepMAB-Classic Oyster 556 1:200 in MF7 cells.

Fixation: 4 % Formaldehyd, 4 °C, 20 min
Kindly provided by: Dr. U. Bertsch, Inst. f. Immunologie, UK Kiel

Strep-NogoB fusion protein stained with StrepMAB-Classic Oyster 488 1:300 in Schwann cells.
Fixation: 100 % MetOH, 10 min, RT
StrepMAB Classic 488 1:300 in 3 % NGS/1 % BSA 4 °C o/n
Kindly provided by: Dr. rer. nat. Rüdiger Schweigreiter, Sektion für Neurobiochemie, Biozentrum, Medizinische Universität Innsbruck

StrepMAB-Immo

FACS

FACS analysis of Zymosan, stained with StrepMAB-Immo Oyster 645 and - Chromeo 642 against Clec7-One-STrEP-tag 1:100, 20 min, on ice. Kindly provided by: Dr. K. Neumann, III Med. Klinik, TUM München

Protein Immobilization

strep-tagII%20_MAK%20Biacore%20Kopie-1880f750.png

During the washing phase, the recombinant Strep-tag® protein of interest remains tightly bound to StrepMAB-Immo, while a significant amount of Strep-tag® protein is washed off using the competitive antibody

Specifications

StrepMAB-Classic

StrepMAB-Classic, Cat. No. 2-1507-001

Purification	purified by affinity chromatography
Host animal/Subclass	Mouse/IgG1
Specificity	N- or C-terminal or internal Strep-tag® II
Form	lyophilized from NH₄HCO₃ with 10 mg sucrose per mg antibod
Stability	18 months after shipment
Storage	stored at -20 °C
Shipment	RT

StrepMAB-Classic, HRP conjugate

StrepMAB-Classic, HRP conjugate, Cat. No. 2-1509-001

Purification	purified by affinity chromatography
HRP conjugate	conjugated to 3-3.5 mols horseradish peroxidase per mol IgG
Host animal/Subclass	Mouse/IgG1
Specificity	N- or C-terminal or internal Strep-tag® II
Form	suspension in PBS with 0.5 % BSA and 0.01 % thimerosal
Stability	18 months after shipment
Storage	stored at 4 °C, do not freeze!
Shipment	RT

StrepMAB-Immo

StrepMAB-Immo, Cat. No. 2-1521-001

Host animal/Subclass	Mouse/IgG1
Specifity	Nearly irreversible binding of C- or N-terminally fused Strep-tag® II, which is N-terminally extended by "SerAla" ** like recombinant protein - SA-WSHPQFEK or SA-WSHPQFEK - recombinant protein. Proteins containing Strep-tag® II with N-terminal sequences other than SerAla are bound with reduced affinity in the cases tested so far.
Form binding capacity	lyophilized from NH₄HCO₃with 10 mg sucrose per mg antibody (determined for bacterial alkaline phosphatase)
Stability	24 months after shipment
Storage	-20 °C
Shipment	RT

**pASK-/pEXPR- and pPR-IBA vectors with N-term One-/Strep-tag® II do NOT have "SerAla"-Linker but the StarGate® vectors (carrying Strep-tag® II and One-STrEP-tag) are equipped with the SA-Linker.

Secondary antibody, HRP conjugate

Rabbit anti Mouse pAb, HRP conjugate Cat. No. 2-1591-001

Description	Purified polyclonal rabbit anti mouse immunoglobulin fraction conjugated to horseradish peroxidase (HRP).
Specifity	The antibody used for conjugation has not been absorbed to remove cross-reactions to immunoglobulin from other species. Therefore the conjugate shows reactions with mouse immunoglobulin of all classes
Form	Dissolved in 50 mM Tris/HCl 15 mM NaN₃, pH 7.2
Concentration of conjugate	1.3 mg/ml
Amount	0.5 ml
Dilution	Dilute 1:1000 for chromogenic detection of bound mouse monoclonal antibodies in Western Blots Dilute 1:2000 (or titrate up to 1:10,000) for detection of mouse monoclonal antibodies in microplate assays
Storage	2 - 8 °C
Stability	6 months after shipping
Shipment	RT

StrepMAB-Classic

Immunofluorescence

Strep-tag-FasL fusion protein stained with StrepMAB-Classic Oyster 556 1:200 in MF7 cells.

Fixation: 4 % Formaldehyd, 4 °C, 20 min
Kindly provided by: Dr. U. Bertsch, Inst. f. Immunologie, UK Kiel

Strep-NogoB fusion protein stained with StrepMAB-Classic Oyster 488 1:300 in Schwann cells.
Fixation: 100 % MetOH, 10 min, RT
StrepMAB Classic 488 1:300 in 3 % NGS/1 % BSA 4 °C o/n
Kindly provided by: Dr. rer. nat. Rüdiger Schweigreiter, Sektion für Neurobiochemie, Biozentrum, Medizinische Universität Innsbruck

StrepMAB-Immo

FACS

Protein Immobilization

References

Seitz P. and Blokesch M. (2013)
DNA-uptake machinery of naturally competent Vibrio cholerae
PNAS 110(44):17987-92. doi: 10.1073/pnas.1315647110
Ernst, W.J., Spenger, A., Toellner, L., Katinger, H. and Grabherr, R.M. (2000)
Expanding baculovirus surface display modification of the native coat protein Gp64 of autographa californica NPV.
Eur. J. Biochem. 267, 4033-4039.
Panke, O., Gumbiowski, K., Junge, W. and Engelbrecht, S. (2000)
F-ATPase: specific observation of the rotating c subunit oligomer of EFoEF1.
FEBS Letters 472, 34-38.
Skerra, A. and Schmidt, T.G.M. (1993)
Use of the strep-tag and streptavidin for detection and purification of recombinant proteins.
Methods in Enzymology 326, 271-304.
Sebastian, P., Wallwitz, J. and Schmidt, S. (2003)
Semi automated production of a set of different recombinant GST-streptag fusion proteins.
J. Chromatogr. B 786, 343-355

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