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Myc tag Rabbit Polyclonal Antibody

Usd: 315

Specification

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Catalog# R1208-1

Myc tag Rabbit Polyclonal Antibody

Application

  • WB

  • IP

  • IF-Cell

  • FC

Reactivity

  • Species independent

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Myc tag Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Synthetic peptide within human Myc aa 410-420.

Species Reactivity

Species independent

Validated Applications

WB, IP, IF-Cell, FC

Positive Control

PG-CM cell lysates, C-terminal Myc-tagged recombinant protein, N-terminal Myc-tagged recombinant protein.

Conjugation

unconjugated

RRID

AB_3073204

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:20,000-1:50,000

  • IP

  • 2-5 µg/ml.

  • IF-Cell

  • 1:200

  • FC

  • 1:1,000

Target

Function

Myc gene encodes for a transcription factor that is believed to regulate expression of 15% of all genes through binding on Enhancer Box sequences (E-boxes) and recruiting histone acetyltransferases (HATs). c-Myc is commonly activated in a variety of tumor cells and plays an important role in cellular proliferation, differentiation, apoptosis and cell cycle progression. This Myc-Tag antibody detects Myc-tagged fusion proteins.

Background References

1. "A quantitative atlas of mitotic phosphorylation."Dephoure N., Zhou C., Villen J., Beausoleil S.A., Bakalarski C.E., Elledge S.J., Gygi S.P.Proc. Natl. Acad. Sci. U.S.A. 105:10762-10767(2008)

2. "Transactivation of gene expression by Myc is inhibited by mutation at the phosphorylation sites Thr-58 and Ser-62."Gupta S., Seth A., Davis R.J. Proc. Natl. Acad. Sci. U.S.A. 90:3216-3220(1993)

Post-translational Modification

Phosphorylated by PRKDC. Phosphorylation at Ser-329 by PIM2 leads to the stabilization of MYC (By similarity). Phosphorylation at Ser-62 by CDK2 prevents Ras-induced senescence. Phosphorylated at Ser-62 by DYRK2; this primes the protein for subsequent phosphorylation by GSK3B at Thr-58. Phosphorylation at Thr-58 and Ser-62 by GSK3 is required for ubiquitination and degradation by the proteasome.; Ubiquitinated by the SCF(FBXW7) complex when phosphorylated at Thr-58 and Ser-62, leading to its degradation by the proteasome. In the nucleoplasm, ubiquitination is counteracted by USP28, which interacts with isoform 1 of FBXW7 (FBW7alpha), leading to its deubiquitination and preventing degradation. In the nucleolus, however, ubiquitination is not counteracted by USP28 but by USP36, due to the lack of interaction between isoform 3 of FBXW7 (FBW7gamma) and USP28, explaining the selective MYC degradation in the nucleolus. Also polyubiquitinated by the DCX(TRUSS) complex. Ubiquitinated by TRIM6 in a phosphorylation-independent manner (By similarity).

UNIPROT

SwissProt: P01106 Human

Synonyms

avian myelocytomatosis viral oncogene homolog antibody

bHLHe39 antibody

c-Myc antibody

class E basic helix-loop-helix protein 39 antibody

MRTL antibody

MYC antibody

Myc Epitope Tag antibody

myc proto-oncogene protein antibody

myc-related translation/localization regulatory factor antibody

oncogene c-Myc antibody

Expand

Images

  • Western blot analysis of Myc tag on different lysates with Rabbit anti-Myc tag antibody (<a href="/products/R1208-1" style="font-weight: bold;text-decoration: underline;">R1208-1</a>) at 1/20,000 dilution.<br /><br />Lane 1: 293T cell lysate<br />Lane 2: 293T transfected with Myc-tagged Claudin18.2 (C-terminal) cell lysate<br />Lane 3: 293T transfected with Myc-tagged Histone H3.1 (N-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/R1208-1" style="font-weight: bold;text-decoration: underline;">R1208-1</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 Myc tag on different lysates with Rabbit anti-Myc tag antibody (R1208-1) at 1/20,000 dilution.

    Lane 1: 293T cell lysate
    Lane 2: 293T transfected with Myc-tagged Claudin18.2 (C-terminal) cell lysate
    Lane 3: 293T transfected with Myc-tagged Histone H3.1 (N-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 (R1208-1) 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.

  • Myc tag was immunoprecipitated in 2µg C terminal Myc Tag fusion protein lysate with <a href="/products/R1208-1" style="font-weight: bold;text-decoration: underline;">R1208-1</a> at 2 µg/20 µl agarose. Western blot was performed from the immunoprecipitate using <a href="/products/EM31105" style="font-weight: bold;text-decoration: underline;">EM31105</a> at 1/1000 dilution. Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:20,000 dilution was used for 60 mins at room temperature.<br /><br />Lane 1: Myc Tag fusion protein lysate (input).<br />Lane 2: <a href="/products/R1208-1" style="font-weight: bold;text-decoration: underline;">R1208-1</a> IP in Myc Tag fusion protein lysate.<br />Lane 3: Rabbit IgG instead of <a href="/products/R1208-1" style="font-weight: bold;text-decoration: underline;">R1208-1</a> in Myc Tag fusion protein lysate.<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST

    Myc tag was immunoprecipitated in 2µg C terminal Myc Tag fusion protein lysate with R1208-1 at 2 µg/20 µl agarose. Western blot was performed from the immunoprecipitate using EM31105 at 1/1000 dilution. Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 60 mins at room temperature.

    Lane 1: Myc Tag fusion protein lysate (input).
    Lane 2: R1208-1 IP in Myc Tag fusion protein lysate.
    Lane 3: Rabbit IgG instead of R1208-1 in Myc Tag fusion protein lysate.

    Blocking/Dilution buffer: 5% NFDM/TBST

  • Myc tag was immunoprecipitated in 2µg N terminal Myc Tag fusion protein lysate with <a href="/products/R1208-1" style="font-weight: bold;text-decoration: underline;">R1208-1</a> at 2 µg/20 µl agarose. Western blot was performed from the immunoprecipitate using <a href="/products/EM31105" style="font-weight: bold;text-decoration: underline;">EM31105</a> at 1/1000 dilution. Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:20,000 dilution was used for 60 mins at room temperature.<br /><br />Lane 1: Myc Tag fusion protein lysate (input).<br />Lane 2: <a href="/products/R1208-1" style="font-weight: bold;text-decoration: underline;">R1208-1</a> IP in Myc Tag fusion protein lysate.<br />Lane 3: Rabbit IgG instead of <a href="/products/R1208-1" style="font-weight: bold;text-decoration: underline;">R1208-1</a> in Myc Tag fusion protein lysate.<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST

    Myc tag was immunoprecipitated in 2µg N terminal Myc Tag fusion protein lysate with R1208-1 at 2 µg/20 µl agarose. Western blot was performed from the immunoprecipitate using EM31105 at 1/1000 dilution. Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 60 mins at room temperature.

    Lane 1: Myc Tag fusion protein lysate (input).
    Lane 2: R1208-1 IP in Myc Tag fusion protein lysate.
    Lane 3: Rabbit IgG instead of R1208-1 in Myc Tag fusion protein lysate.

    Blocking/Dilution buffer: 5% NFDM/TBST

  • Immunocytochemistry analysis of 293T cells labeling Myc tag with Rabbit anti-Myc tag antibody (<a href="/products/R1208-1" style="font-weight: bold;text-decoration: underline;">R1208-1</a>) at 1/200 dilution.<br /><br />293T cells, transfected with Myc-tagged empty control, Claudin18.2 (C-terminal) or Histone H3.1 (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-Myc tag antibody (<a href="/products/R1208-1" style="font-weight: bold;text-decoration: underline;">R1208-1</a>) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</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 />Myc Tag (<a href="/products/HA601081" style="font-weight: bold;text-decoration: underline;">HA601081</a>, green) was stained at 1/1,000 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of 293T cells labeling Myc tag with Rabbit anti-Myc tag antibody (R1208-1) at 1/200 dilution.

    293T cells, transfected with Myc-tagged empty control, Claudin18.2 (C-terminal) or Histone H3.1 (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-Myc tag antibody (R1208-1) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) 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.

    Myc Tag (HA601081, green) was stained at 1/1,000 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of 293T cells transfected C-myc-tag labeling Myc tag.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/R1208-1" style="font-weight: bold;text-decoration: underline;">R1208-1</a>, 1μg/mL) (red) compared with Rabbit IgG 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-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</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 293T cells transfected C-myc-tag labeling Myc tag.

    Cells were fixed and permeabilized. Then stained with the primary antibody (R1208-1, 1μg/mL) (red) compared with Rabbit IgG 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-Rabbit IgG Secondary antibody (HA1121) 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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Citation

Products with the same target and pathway

metafield image

HRP Conjugated Myc tag Rabbit Polyclonal Antibody

Application: WB

Reactivity: Species independent

Conjugate: HRP