Strep-tagŪII-Tag antibodies (polyclonal)
AS21 4527 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Strep-tag®II-Taged proteins

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Product Information
Immunogen
KLH-conjugated synthetic peptide Strep-tag®II epitope tag, sequence: ASWSHPQFEKGA
Host
Rabbit
Clonality
Polyclonal
Purity
Immunogen affinity purified serum, in PBS pH 7.4.
Format
Lyophilized
Quantity
50 ĩg
Reconstitution
For reconstitution add 50 µl, of sterile water.
Storage
Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Tested applications
Western blot (WB)
Recommended dilution
1 : 1000 (WB)
Reactivity
Confirmed reactivity
Strep-tag®II-proteins
Application examples
Application examples

Soluble (4 µg protein) and membrane proteins (corresponding to 0,25 µg of chlorophyll a) from Synechocystis sp. PCC 6803 were extracted with and resuspended in ACA buffer (750 mM -amino caproic acid; 50 mM BisTris/HCl, pH 7.0; 0.5 mM EDTA). Samples were denatured with 2x sample buffer (125mM Tris, pH=6,8; 200mM DTT; 4% (w/v) SDS; 20% (w/v) Glycerin; 0,02% (w/v) bromophenolblue) at room temperature (RT) for 1h. The proteins were separated on 12.5 % SDS PAGE (Bis-Tris) gels and blotted for 60 min onto a nitrocellulose membrane using a wet transfer system (BioRad). The membrane was blocked with 5% milk powder in PBS-T for 1 h at RT with agitation. The blot was then incubated overnight with the primary antibody (anti Strep-tag, Agrisera, AS21 4527) at a dilution of 1:1.000 in PBS-T at 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly, then washed three times for 20 min in PBS-T with agitation. The blot was incubated using a matching secondary antibody (anti-rabbit IgG horseradish peroxidase conjugated) diluted to 1:10 000 in PBS-T for 1 h at RT with agitation. The blot was washed three times for 10 min with PBS-T and two times for 10 minutes with PBS. Subsequently, the membrane was incubated with AgriseraECL SuperBright solutions (AS16 ECL-S-10) for 1 minute. The differently exposed films were developed using a NDT DÜRR developer.

Soluble (4 µg protein) and membrane proteins (corresponding to 0,25 µg of chlorophyll a) from Synechocystis sp. PCC 6803 were extracted with and resuspended in ACA buffer (750 mM -amino caproic acid; 50 mM BisTris/HCl, pH 7.0; 0.5 mM EDTA). Samples were denatured with 2x sample buffer (125mM Tris, pH=6,8; 200mM DTT; 4% (w/v) SDS; 20% (w/v) Glycerin; 0,02% (w/v) bromophenolblue) at room temperature (RT) for 1h. The proteins were separated on 12.5 % SDS PAGE (Bis-Tris) gels and blotted for 60 min onto a nitrocellulose membrane using a wet transfer system (BioRad). The membrane was blocked with 5% milk powder in PBS-T for 1 h at RT with agitation. The blot was then incubated overnight with the primary antibody (anti Strep-tag, Agrisera, AS21 4527) at a dilution of 1:1.000 in PBS-T at 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly, then washed three times for 20 min in PBS-T with agitation. The blot was incubated using a matching secondary antibody (anti-rabbit IgG horseradish peroxidase conjugated) diluted to 1:10 000 in PBS-T for 1 h at RT with agitation. The blot was washed three times for 10 min with PBS-T and two times for 10 minutes with PBS. Subsequently, the membrane was incubated with AgriseraECL SuperBright solutions (AS16 ECL-S-10) for 1 minute. The differently exposed films were developed using a NDT DÜRR developer.
Courtesy of Dr. Marko Böhm, University of Kiel, Germany
Additional information
Background
Background
Strep-tag®II is a synthetic peptide consisting of eight amino acids (Trp-Ser-His-Pro-Gln-Phe-Glu-Lys) and this peptide sequence shows high affinity towards Strep-Tactin®, a specifically engineered streptavidin, and can be N- or C- terminally fused to recombinant proteins.
Product citations
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