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Strep-Tag II Recombinant Rabbit Monoclonal Antibody [PSH05-13] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750986

Strep-Tag II Recombinant Rabbit Monoclonal Antibody [PSH05-13] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • IP

Reactivity

  • Species independent

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

Strep-Tag II Recombinant Rabbit Monoclonal Antibody [PSH05-13] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide corresponding to Strep-tag II.

Species Reactivity

Species independent

Validated Applications

WB, IF-Cell, IP

Conjugation

unconjugated

Clone Number

PSH05-13

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:10,000

  • IF-Cell

  • 1:2,500

  • IP

  • 1-2μg/sample

Target

Function

The Strep-tag® system is a method which allows the purification and detection of proteins by affinity chromatography. The Strep-tag II is a synthetic peptide consisting of eight amino acids (Trp-Ser-His-Pro-Gln-Phe-Glu-Lys). This peptide sequence exhibits intrinsic affinity towards Strep-Tactin®, a specifically engineered streptavidin, and can be N- or C- terminally fused to recombinant proteins. By exploiting the highly specific interaction, Strep-tagged proteins can be isolated in one step from crude cell lysates. Because the Strep-tag elutes under gentle, physiological conditions it is especially suited for generation of functional proteins.

Background References

1. Arne Skerra. et. al. The Strep-tag system for one-step purification and high-affinity detection or capturing of proteins. Nat Protoc. 2007;2(6):1528-35.

Synonyms

anti-Strep-Tag II antibody

Strep-Tag II antibody

Strep-Tag II antibody antibody

Strep-Tag IIantiboy antibody

Images

  • Western blot analysis of Strep-Tag II on different lysates with Rabbit anti-Strep-Tag II antibody (<a href="/products/HA750986" style="font-weight: bold;text-decoration: underline;">HA750986</a>) at 1/10,000 dilution.<br /><br />Lane 1: 293T transfected with Strep-Tag II-tagged Histone H3.1 (C-terminal) cell lysate<br />Lane 2: 293T transfected with Strep-Tag II-tagged ACAT2 (N-terminal) cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Exposure time: 2 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750986" style="font-weight: bold;text-decoration: underline;">HA750986</a>) at 1/10,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Strep-Tag II on different lysates with Rabbit anti-Strep-Tag II antibody (HA750986) at 1/10,000 dilution.

    Lane 1: 293T transfected with Strep-Tag II-tagged Histone H3.1 (C-terminal) cell lysate
    Lane 2: 293T transfected with Strep-Tag II-tagged ACAT2 (N-terminal) cell lysate

    Lysates/proteins at 10 µg/Lane.

    Exposure time: 2 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750986) at 1/10,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling Strep-Tag II with Rabbit anti-Strep-Tag II antibody (<a href="/products/HA750986" style="font-weight: bold;text-decoration: underline;">HA750986</a>) at 1/2,500 dilution.<br /><br />HeLa cells, transfected with Strep-Tag II-tagged Histone H3.1 (C-terminal) or ACAT2 (N-terminal) expression vector, respectively, were fixed in 4% paraformaldehyde for 10 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Strep-Tag II antibody (<a href="/products/HA750986" style="font-weight: bold;text-decoration: underline;">HA750986</a>) at 1/2,500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling Strep-Tag II with Rabbit anti-Strep-Tag II antibody (HA750986) at 1/2,500 dilution.

    HeLa cells, transfected with Strep-Tag II-tagged Histone H3.1 (C-terminal) or ACAT2 (N-terminal) expression vector, respectively, were fixed in 4% paraformaldehyde for 10 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Strep-Tag II antibody (HA750986) at 1/2,500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Strep-Tag II was immunoprecipitated from 0.2 mg 293T transfected with Strep-Tag II-tagged Histone H3.1 (C-terminal) cell lysate with <a href="/products/HA750986" style="font-weight: bold;text-decoration: underline;">HA750986</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA750986" style="font-weight: bold;text-decoration: underline;">HA750986</a> at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: 293T transfected with Strep-Tag II-tagged Histone H3.1 (C-terminal) cell lysate (input)<br />Lane 2: <a href="/products/HA750986" style="font-weight: bold;text-decoration: underline;">HA750986</a> IP in 293T transfected with Strep-Tag II-tagged Histone H3.1 (C-terminal) cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA750986" style="font-weight: bold;text-decoration: underline;">HA750986</a> in 293T transfected with Strep-Tag II-tagged Histone H3.1 (C-terminal) cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 2 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    Strep-Tag II was immunoprecipitated from 0.2 mg 293T transfected with Strep-Tag II-tagged Histone H3.1 (C-terminal) cell lysate with HA750986 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA750986 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: 293T transfected with Strep-Tag II-tagged Histone H3.1 (C-terminal) cell lysate (input)
    Lane 2: HA750986 IP in 293T transfected with Strep-Tag II-tagged Histone H3.1 (C-terminal) cell lysate
    Lane 3: Rabbit IgG instead of HA750986 in 293T transfected with Strep-Tag II-tagged Histone H3.1 (C-terminal) cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 2 seconds; ECL: K1801

  • Strep-Tag II was immunoprecipitated from 0.2 mg 293T transfected with Strep-Tag II-tagged ACAT2 (N-terminal) cell lysate with <a href="/products/HA750986" style="font-weight: bold;text-decoration: underline;">HA750986</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA750986" style="font-weight: bold;text-decoration: underline;">HA750986</a> at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: 293T transfected with Strep-Tag II-tagged ACAT2 (N-terminal) cell lysate (input)<br />Lane 2: <a href="/products/HA750986" style="font-weight: bold;text-decoration: underline;">HA750986</a> IP in 293T transfected with Strep-Tag II-tagged ACAT2 (N-terminal) cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA750986" style="font-weight: bold;text-decoration: underline;">HA750986</a> in 293T transfected with Strep-Tag II-tagged ACAT2 (N-terminal) cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 2 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    Strep-Tag II was immunoprecipitated from 0.2 mg 293T transfected with Strep-Tag II-tagged ACAT2 (N-terminal) cell lysate with HA750986 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA750986 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: 293T transfected with Strep-Tag II-tagged ACAT2 (N-terminal) cell lysate (input)
    Lane 2: HA750986 IP in 293T transfected with Strep-Tag II-tagged ACAT2 (N-terminal) cell lysate
    Lane 3: Rabbit IgG instead of HA750986 in 293T transfected with Strep-Tag II-tagged ACAT2 (N-terminal) cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 2 seconds; ECL: K1801

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