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Strep-tagŪII-Tag antibodies (polyclonal)

AS21 4527   | Clonality: Polyclonal |  Host: Rabbit  | Reactivity: Strep-tag®II-Taged proteins

Strep-tagŪII-Tag antibodies (polyclonal) in the group Tag Antibodies / Halo/HSV/KT3/Strep at Agrisera AB (Antibodies for research) (AS21 4527)
Strep-tagŪII-Tag antibodies (polyclonal)



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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
Western blot using anti-Strep tag antibodies (polyclonal)
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

Related products

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

immunogen: KLH-conjugated synthetic peptide Strep-tag®II epitope tag, sequence: ASWSHPQFEKGA
Reconstitution: For reconstitution add 50 µl, of sterile water.
Host: Rabbit
Clonality: Polyclonal
Purity: Immunogen affinity purified serum, in PBS pH 7.4.
Format: Lyophilized
Quantity: 50 ĩg
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)
Confirmed reactivity: Strep-tag®II-proteins
Picture (footer):
Western blot using anti-Strep tag antibodies (polyclonal)
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

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.

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