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HA tag Recombinant Rabbit Multiclonal Antibody [PSH01-92] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750766

HA tag Recombinant Rabbit Multiclonal Antibody [PSH01-92] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • IP

Reactivity

  • Species independent

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

HA tag Recombinant Rabbit Multiclonal Antibody [PSH01-92] - BSA and Azide free

Antibody Type

Recombinant Rabbit multiclonal

Immunogen

Synthetic peptide within N Terminal fusion protein and C Terminal fusion protein.

Species Reactivity

Species independent

Validated Applications

WB, IF-Cell, IP

Positive Control

N Terminal fusion protein and C Terminal fusion protein.

Conjugation

unconjugated

Clone Number

PSH01-92

Product Features

Form

Liquid

Concentration

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

  • 1:5,000

  • IF-Cell

  • 1:1,000

  • IP

  • 1-2μg/sample

Target

Function

Human influenza hemagglutinin (HA) is a surface glycoprotein required for the infectivity of the human virus. The HA tag is derived from the HA molecule corresponding to amino acids 98-106 has been extensively used as a general epitope tag in expression vectors. Many recombinant proteins have been engineered to express the HA tag, which does not appear to interfere with the bioactivity or the biodistribution of the recombinant protein. This tag facilitates the detection, isolation, and purification of the proteins.

Background References

1. Liu Z et al. Mark4 promotes oxidative stress and inflammation via binding to PPAR and activating NF-kB pathway in mice adipocytes. Sci Rep 6:21382 (2016).

2. Gauson EJ et al. Evidence supporting a role for TopBP1 and Brd4 in the initiation but not continuation of human papillomavirus 16 E1/E2-mediated DNA replication. J Virol 89:4980-91 (2015).

Synonyms

HA epitope tag antibody

HA1 antibody

HA2 antibody

hemagglutinin antibody

Hemagglutinin HA1 chain antibody

Hemagglutinin HA2 chain antibody

Images

  • Western blot analysis of HA tag on different lysates with Rabbit anti-HA tag antibody (<a href="/products/HA750766" style="font-weight: bold;text-decoration: underline;">HA750766</a>) at 1/5,000 dilution.<br /><br />Lane 1: 293T cell lysate<br />Lane 2: 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate<br />Lane 3: 293T transfected with HA-tagged LIPT1 (N-terminal) cell lysate<br />Lane 4: 293T transfected with HA-tagged CXCL13 (C-terminal) cell lysate<br />Lane 5: 293T transfected with HA-tagged CLK4 (C-terminal) cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Exposure time: 3 minutes;<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/HA750766" style="font-weight: bold;text-decoration: underline;">HA750766</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. 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 HA tag on different lysates with Rabbit anti-HA tag antibody (HA750766) at 1/5,000 dilution.

    Lane 1: 293T cell lysate
    Lane 2: 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate
    Lane 3: 293T transfected with HA-tagged LIPT1 (N-terminal) cell lysate
    Lane 4: 293T transfected with HA-tagged CXCL13 (C-terminal) cell lysate
    Lane 5: 293T transfected with HA-tagged CLK4 (C-terminal) cell lysate

    Lysates/proteins at 10 µg/Lane.

    Exposure time: 3 minutes;

    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 (HA750766) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of 293T cells labeling HA tag with Rabbit anti-HA tag antibody (<a href="/products/HA750766" style="font-weight: bold;text-decoration: underline;">HA750766</a>) at 1/1,000 dilution.<br /><br />293T cells, transfected with HA-tagged empty control, Nanos homolog 3 (N-terminal) or CLK4 (C-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-HA tag antibody (<a href="/products/HA750766" style="font-weight: bold;text-decoration: underline;">HA750766</a>) at 1/1,000 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 />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 293T cells labeling HA tag with Rabbit anti-HA tag antibody (HA750766) at 1/1,000 dilution.

    293T cells, transfected with HA-tagged empty control, Nanos homolog 3 (N-terminal) or CLK4 (C-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-HA tag antibody (HA750766) at 1/1,000 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.

    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.

  • <span style="font-weight: bold;">☑ Independent antibody validation (IAV)</span><br /><br />HA tag was immunoprecipitated from 0.2 mg 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate with <a href="/products/HA750766" style="font-weight: bold;text-decoration: underline;">HA750766</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/M1008-1" style="font-weight: bold;text-decoration: underline;">M1008-1</a> at 1/20,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (<a href="/products/NBI02H" style="font-weight: bold;text-decoration: underline;">NBI02H</a>) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate (input)<br />Lane 2: <a href="/products/HA750766" style="font-weight: bold;text-decoration: underline;">HA750766</a> IP in 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate<br />Lane 3: Mouse IgG instead of <a href="/products/HA750766" style="font-weight: bold;text-decoration: underline;">HA750766</a> in 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 35 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    ☑ Independent antibody validation (IAV)

    HA tag was immunoprecipitated from 0.2 mg 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate with HA750766 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using M1008-1 at 1/20,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate (input)
    Lane 2: HA750766 IP in 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate
    Lane 3: Mouse IgG instead of HA750766 in 293T transfected with HA-tagged Nanos homolog 3 (N-terminal) cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 35 seconds; ECL: K1801

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