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V5 tag Recombinant Rabbit Monoclonal Antibody [SY30-01] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750086

V5 tag Recombinant Rabbit Monoclonal Antibody [SY30-01] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • IP

Reactivity

  • Species independent

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

V5 tag Recombinant Rabbit Monoclonal Antibody [SY30-01] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide immune sequence is CGKPIPNPLLGLDST.

Species Reactivity

Species independent

Validated Applications

WB, IF-Cell, IP

Positive Control

V5 tagged recombinant protein.

Conjugation

unconjugated

Clone Number

SY30-01

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:20,000-1:50,000

  • IF-Cell

  • 1:2,000

  • IP

  • 1-2μg/sample

Target

Function

The V5 epitope tag is derived from a small epitope (Pk) present on the P and V proteins of the paramyxovirus of simian virus 5 (SV5). The anti-V5 antibody detects recombinant proteins containing the 14 amino acid GKPIPNPLLGLDST sequence.

Background References

1. Tatenhorst L et al. Fasudil attenuates aggregation of a-synuclein in models of Parkinson\'s disease. Acta Neuropathol Commun 4:39 (2016).

2. Ji H et al. EGFR phosphorylates FAM129B to promote Ras activation. Proc Natl Acad Sci U S A 113:644-9 (2016).

Synonyms

GKPIPNPLLGLDST epitope tag antibody

GKPIPNPLLGLDST tag antibody

Protein Rev antibody

Regulator of expression of viral proteins antibody

rev antibody

V5 epitope tag antibody

Images

  • Western blot analysis of V5 tag on different lysates with Rabbit anti-V5 tag antibody (<a href="/products/HA750086" style="font-weight: bold;text-decoration: underline;">HA750086</a>) at 1/20,000 dilution.<br /><br />Lane 1: 293T cell lysate<br />Lane 2: 293T transfected with V5-tagged Histone H3.1 (N-terminal) cell lysate<br />Lane 3: 293T transfected with V5-tagged Nanos homolog 3 (C-terminal) cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Exposure time: 2 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/HA750086" style="font-weight: bold;text-decoration: underline;">HA750086</a>) at 1/20,000 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 V5 tag on different lysates with Rabbit anti-V5 tag antibody (HA750086) at 1/20,000 dilution.

    Lane 1: 293T cell lysate
    Lane 2: 293T transfected with V5-tagged Histone H3.1 (N-terminal) cell lysate
    Lane 3: 293T transfected with V5-tagged Nanos homolog 3 (C-terminal) cell lysate

    Lysates/proteins at 10 µg/Lane.

    Exposure time: 2 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 (HA750086) at 1/20,000 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 V5 tag with Rabbit anti-V5 tag antibody (<a href="/products/HA750086" style="font-weight: bold;text-decoration: underline;">HA750086</a>) at 1/2,000 dilution.<br /><br />HeLa cells, transfected with V5-tagged empty control, Histone H3 (N-terminal) or Nanos homolog 3 (C-terminal) expression vector, respectively, were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-V5 tag antibody (<a href="/products/HA750086" style="font-weight: bold;text-decoration: underline;">HA750086</a>) at 1/2,000 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 />V5 tag (<a href="/products/M1008-2" style="font-weight: bold;text-decoration: underline;">M1008-2</a>, red) was stained at 1/1,000 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 V5 tag with Rabbit anti-V5 tag antibody (HA750086) at 1/2,000 dilution.

    HeLa cells, transfected with V5-tagged empty control, Histone H3 (N-terminal) or Nanos homolog 3 (C-terminal) expression vector, respectively, were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-V5 tag antibody (HA750086) at 1/2,000 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.

    V5 tag (M1008-2, red) was stained at 1/1,000 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • V5 tag was immunoprecipitated from 0.2 mg 293T transfected with V5-tagged Histone H3.1 (N-terminal) cell lysate with <a href="/products/HA750086" style="font-weight: bold;text-decoration: underline;">HA750086</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA750086" style="font-weight: bold;text-decoration: underline;">HA750086</a> at 1/10,000 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 V5-tagged Histone H3.1 (N-terminal) cell lysate (input)<br />Lane 2: <a href="/products/HA750086" style="font-weight: bold;text-decoration: underline;">HA750086</a> IP in 293T transfected with V5-tagged Histone H3.1 (N-terminal) cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA750086" style="font-weight: bold;text-decoration: underline;">HA750086</a> in 293T transfected with V5-tagged Histone H3.1 (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>

    V5 tag was immunoprecipitated from 0.2 mg 293T transfected with V5-tagged Histone H3.1 (N-terminal) cell lysate with HA750086 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA750086 at 1/10,000 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 V5-tagged Histone H3.1 (N-terminal) cell lysate (input)
    Lane 2: HA750086 IP in 293T transfected with V5-tagged Histone H3.1 (N-terminal) cell lysate
    Lane 3: Rabbit IgG instead of HA750086 in 293T transfected with V5-tagged Histone H3.1 (N-terminal) cell lysate

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

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"