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Strep-Tag II Mouse Monoclonal Antibody [A5D11]

Usd: 350

Specification

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Catalog# HA600038

Strep-Tag II Mouse Monoclonal Antibody [A5D11]

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Species independent

Conjugation

  • unconjugated

Overview

Product Name

Strep-Tag II Mouse Monoclonal Antibody [A5D11]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Synthetic peptide corresponding to Strep-tag II.

Species Reactivity

Species independent

Validated Applications

WB, IF-Cell, FC

Conjugation

unconjugated

Clone Number

A5D11

RRID

AB_3071655

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein G affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:1,000

  • FC

  • 1:1,000

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 Mouse anti-Strep-Tag II antibody (<a href="/products/HA600038" style="font-weight: bold;text-decoration: underline;">HA600038</a>) at 1/1,000 dilution.<br /><br />Lane 1: 293T-NT cell lysate<br />Lane 2: 293T transfected with Strep-Tag II-tagged Histone H3.1 (C-terminal) cell lysate<br />Lane 3: 293T transfected with Strep-Tag II-tagged ACAT2 (N-terminal) cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Exposure time: 30 seconds; ECL: K1801;<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/HA600038" style="font-weight: bold;text-decoration: underline;">HA600038</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</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 Mouse anti-Strep-Tag II antibody (HA600038) at 1/1,000 dilution.

    Lane 1: 293T-NT cell lysate
    Lane 2: 293T transfected with Strep-Tag II-tagged Histone H3.1 (C-terminal) cell lysate
    Lane 3: 293T transfected with Strep-Tag II-tagged ACAT2 (N-terminal) cell lysate

    Lysates/proteins at 20 µg/Lane.

    Exposure time: 30 seconds; ECL: K1801;

    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 (HA600038) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling Strep-Tag II with Mouse anti-Strep-Tag II antibody (<a href="/products/HA600038" style="font-weight: bold;text-decoration: underline;">HA600038</a>) at 1/1,000 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 Mouse anti-Strep-Tag II antibody (<a href="/products/HA600038" style="font-weight: bold;text-decoration: underline;">HA600038</a>) at 1/1,000 dilution in 1% BSA in PBST 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. 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/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, green) was stained at 1/100 dilution 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>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling Strep-Tag II with Mouse anti-Strep-Tag II antibody (HA600038) at 1/1,000 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 Mouse anti-Strep-Tag II antibody (HA600038) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) 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 (ET1602-4, green) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) were used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of HeLa cells labeling Strep-Tag II.<br /><br />HeLa cells, transfected with Strep-Tag II-tagged empty control or ACAT2 (N-terminal) expression vector, respectively, were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA600038" style="font-weight: bold;text-decoration: underline;">HA600038</a>, 1μg/mL) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Mouse IgG Secondary antibody (<a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling Strep-Tag II.

    HeLa cells, transfected with Strep-Tag II-tagged empty control or ACAT2 (N-terminal) expression vector, respectively, were fixed and permeabilized. Then stained with the primary antibody (HA600038, 1μg/mL) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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