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ERK-MAPK Pathway Antibody Kit

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Catalog# HAK21117

ERK-MAPK Pathway Antibody Kit

Application

  • WB

Reactivity

  • Human

  • Mouse

Overview

Kit Components

Product Includes Specification Application Reactivity Mw
Phospho-MEK1 (T292)[ET1612-42] 20µl WB,IHC-P Human,Mouse,Rat Predicted band size: 43 kDa
Phospho-Erk1 (T202 + Y204) + Erk2 (T185 + Y187)[ET1610-13] 20µl WB,IHC-P,IF-Cell,IF-Tissue,FC,IP Human,Mouse,Rat Predicted band size: 41/43 kDa
Phospho-RSK1 (S380)[ET1610-28] 20µl WB,IF-Cell,IF-Tissue,IP Human,Mouse,Rat Predicted band size: 83 kDa
Phospho-c-Jun (S73)+JunD (S100)[HA722475] 20µl WB,IF-Cell,FC Human,Mouse,Rat Predicted band size: 40/45 kDa
Phospho-MNK1 (T250 + T255)[HA723960] 20µl WB,IHC-P,IF-Cell,FC Human,Mouse Predicted band size: 51 kDa
Goat Anti-Rabbit IgG (H+L)[HA1001] 100µl WB,ELISA,IHC-P Rabbit

Product Description

The ERK-MAPK Pathway Antibody Kit provides an economical means to investigate the relative levels of key phosphorylated proteins involved at various stages of the ERK-MAPK pathway. The ERK-MAPK Pathway Antibody Kit contains enough primary and secondary antibodies to perform two Western blot experiments.

Product Features

Storage Buffer

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

Storage Instructions

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

Background

This carefully selected series of antibodies provides comprehensive coverage of key phosphorylation events in the ERK-MAPK signaling pathway, making it an ideal tool for studying this cascade. The ERK-MAPK signaling pathway regulates various physiological activities, including cell proliferation, differentiation, and survival, in response to extracellular signals such as growth factors, hormonal stimuli, and environmental stress. <br>This pathway involves a cascade of reactions where protein kinases sequentially phosphorylate each other, leading to the activation of downstream gene expression. Phospho-MEK1 is a MAP2K that, when phosphorylated at multiple sites (including Thr202/Tyr204), activates ERK1/2. Once activated, phosphorylated ERK translocates into the nucleus and phosphorylates several downstream transcription factors.

Data Links

Background References

1. Guo YJ, Pan WW, Liu SB, Shen ZF, Xu Y, Hu LL. ERK/MAPK signalling pathway and tumorigenesis. Exp Ther Med. 2020 Mar;19(3):1997-2007.

2. Iroegbu JD, Ijomone OK, Femi-Akinlosotu OM, Ijomone OM. ERK/MAPK signalling in the developing brain: Perturbations and consequences. Neurosci Biobehav Rev. 2021

3. Fang JY, Richardson BC. The MAPK signalling pathways and colorectal cancer. Lancet Oncol. 2005 May;6(5):322-7.

Images

  • Western blot analysis of Phospho-MEK1 (T292) on different lysates with Rabbit anti-Phospho-MEK1 (T292) antibody (<a href="/products/ET1612-42" style="font-weight: bold;text-decoration: underline;">ET1612-42</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 100ng/mL Nocodazole for 17 hours cell lysate<br />Lane 3: PC-12 cell lysate<br />Lane 4: PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate<br />Lane 5: HeLa treated with 100ng/mL Nocodazole for 17 hours cell lysate, then the membrane treated with λpp for 1 hour<br />Lane 6: PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate, then the membrane treated with λpp for 1 hour<br />Lane 7: NIH/3T3 cell lysate<br />Lane 8: NIH/3T3 treated with 100ng/mL Nocodazole for 18 hours cell lysate<br />Lane 9: NIH/3T3 treated with 100ng/mL Nocodazole for 18 hours cell lysate, then the membrane treated with λpp for 1 hour<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 43 kDa<br />Observed band size: 43 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<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/ET1612-42" style="font-weight: bold;text-decoration: underline;">ET1612-42</a>) at 1/1,000 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 Phospho-MEK1 (T292) on different lysates with Rabbit anti-Phospho-MEK1 (T292) antibody (ET1612-42) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with 100ng/mL Nocodazole for 17 hours cell lysate
    Lane 3: PC-12 cell lysate
    Lane 4: PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate
    Lane 5: HeLa treated with 100ng/mL Nocodazole for 17 hours cell lysate, then the membrane treated with λpp for 1 hour
    Lane 6: PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate, then the membrane treated with λpp for 1 hour
    Lane 7: NIH/3T3 cell lysate
    Lane 8: NIH/3T3 treated with 100ng/mL Nocodazole for 18 hours cell lysate
    Lane 9: NIH/3T3 treated with 100ng/mL Nocodazole for 18 hours cell lysate, then the membrane treated with λpp for 1 hour

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 43 kDa
    Observed band size: 43 kDa

    Exposure time: 3 minutes; 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 (ET1612-42) at 1/1,000 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.

  • Flow cytometric analysis of NIH/3T3 cells untreated (left) / treated with λpp (right) labeling Phospho-MNK1 (T250 + T255).<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA723960" style="font-weight: bold;text-decoration: underline;">HA723960</a>, 1/1,000) (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 NIH/3T3 cells untreated (left) / treated with λpp (right) labeling Phospho-MNK1 (T250 + T255).

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA723960, 1/1,000) (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).

  • Immunocytochemistry analysis of HeLa cells treated with UV for 1 hour labeling Phospho-c-Jun (S73)+JunD (S100) with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (<a href="/products/HA722800" style="font-weight: bold;text-decoration: underline;">HA722800</a>) at 1/500 dilution.<br /><br />Cells 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-Phospho-c-Jun (S73)+JunD (S100) antibody (<a href="/products/HA722800" style="font-weight: bold;text-decoration: underline;">HA722800</a>) at 1/500 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 treated with UV for 1 hour labeling Phospho-c-Jun (S73)+JunD (S100) with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (HA722800) at 1/500 dilution.

    Cells 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-Phospho-c-Jun (S73)+JunD (S100) antibody (HA722800) at 1/500 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.

  • Western blot analysis of Phospho-c-Jun (S73)+JunD (S100) on different lysates with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (<a href="/products/HA722800" style="font-weight: bold;text-decoration: underline;">HA722800</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: HeLa treated with 25μg/mL Anisomycin for 30 minutes cell lysate (20 µg/Lane)<br />Lane 3: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 4: NIH/3T3 treated with 250ng/mL Anisomycin for 30 minutes cell lysate (20 µg/Lane)<br />Lane 5: C6 cell lysate (20 µg/Lane)<br />Lane 6: C6 treated with 25μg/mL Anisomycin for 30 minutes cell lysate (20 µg/Lane)<br /><br />Predicted band size: 36 kDa<br />Observed band size: 36-40 kDa<br /><br />Exposure time: 30 seconds; ECL: K1801;<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/HA722800" style="font-weight: bold;text-decoration: underline;">HA722800</a>) at 1/2,000 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 Phospho-c-Jun (S73)+JunD (S100) on different lysates with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (HA722800) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: HeLa treated with 25μg/mL Anisomycin for 30 minutes cell lysate (20 µg/Lane)
    Lane 3: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 4: NIH/3T3 treated with 250ng/mL Anisomycin for 30 minutes cell lysate (20 µg/Lane)
    Lane 5: C6 cell lysate (20 µg/Lane)
    Lane 6: C6 treated with 25μg/mL Anisomycin for 30 minutes cell lysate (20 µg/Lane)

    Predicted band size: 36 kDa
    Observed band size: 36-40 kDa

    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 (HA722800) at 1/2,000 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.

  • Western blot analysis of Phospho-RSK1 (S380) on different lysates with Rabbit anti-Phospho-RSK1 (S380) antibody (<a href="/products/ET1610-28" style="font-weight: bold;text-decoration: underline;">ET1610-28</a>) at 1/500 dilution.<br /><br />Lane 1: Hela whole cell lysate<br />Lane 2: Hela (starved overnight) then treated with TPA (200nM 4h) cell lysate<br />Lane 3: NIH/3T3 whole cell lysate<br />Lane 4: NIH/3T3 (starved 24h) then Calyculin A (100nM 15min) cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 83 kDa<br />Observed band size: 83 kDa<br /><br />Exposure time: 1 minute;<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/ET1610-28" style="font-weight: bold;text-decoration: underline;">ET1610-28</a>) at 1/500 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 1:200,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Phospho-RSK1 (S380) on different lysates with Rabbit anti-Phospho-RSK1 (S380) antibody (ET1610-28) at 1/500 dilution.

    Lane 1: Hela whole cell lysate
    Lane 2: Hela (starved overnight) then treated with TPA (200nM 4h) cell lysate
    Lane 3: NIH/3T3 whole cell lysate
    Lane 4: NIH/3T3 (starved 24h) then Calyculin A (100nM 15min) cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 83 kDa
    Observed band size: 83 kDa

    Exposure time: 1 minute;

    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 (ET1610-28) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

  • Flow cytometric analysis of HeLa cells untreated (left) or treated (right) with PMA for 30 minutes labeling Phospho-Erk1 (T202 + Y204) + Erk2 (T185 + Y187).<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1610-13" style="font-weight: bold;text-decoration: underline;">ET1610-13</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 HeLa cells untreated (left) or treated (right) with PMA for 30 minutes labeling Phospho-Erk1 (T202 + Y204) + Erk2 (T185 + Y187).

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1610-13, 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).

  • Western blot analysis of Phospho-MNK1 (T250 + T255) on different lysates with Rabbit anti-Phospho-MNK1 (T250 + T255) antibody (<a href="/products/HA723960" style="font-weight: bold;text-decoration: underline;">HA723960</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate<br />Lane 3: HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour<br />Lane 4: NIH/3T3 starved for 24 hours cell lysate<br />Lane 5: NIH/3T3 starved for 24 hours then treated with 10% FBS for 30 minutes cell lysate<br />Lane 6: NIH/3T3 starved for 24 hours then treated with 10% FBS for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 51 kDa<br />Observed band size: 45 kDa<br /><br />Exposure time: 1 minute 16 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/HA723960" style="font-weight: bold;text-decoration: underline;">HA723960</a>) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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 Phospho-MNK1 (T250 + T255) on different lysates with Rabbit anti-Phospho-MNK1 (T250 + T255) antibody (HA723960) at 1/5,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate
    Lane 3: HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour
    Lane 4: NIH/3T3 starved for 24 hours cell lysate
    Lane 5: NIH/3T3 starved for 24 hours then treated with 10% FBS for 30 minutes cell lysate
    Lane 6: NIH/3T3 starved for 24 hours then treated with 10% FBS for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 51 kDa
    Observed band size: 45 kDa

    Exposure time: 1 minute 16 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 (HA723960) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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 NIH/3T3 cells treated with 200nM PMA for 30 minutes labeling Phospho-Erk1 (T202 + Y204) + Erk2 (T185 + Y187) with Rabbit anti-Phospho-Erk1 (T202 + Y204) + Erk2 (T185 + Y187) antibody (<a href="/products/ET1610-13" style="font-weight: bold;text-decoration: underline;">ET1610-13</a>) at 1/100 dilution.<br /><br />Cells 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-Phospho-Erk1 (T202 + Y204) + Erk2 (T185 + Y187) antibody (<a href="/products/ET1610-13" style="font-weight: bold;text-decoration: underline;">ET1610-13</a>) at 1/100 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 NIH/3T3 cells treated with 200nM PMA for 30 minutes labeling Phospho-Erk1 (T202 + Y204) + Erk2 (T185 + Y187) with Rabbit anti-Phospho-Erk1 (T202 + Y204) + Erk2 (T185 + Y187) antibody (ET1610-13) at 1/100 dilution.

    Cells 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-Phospho-Erk1 (T202 + Y204) + Erk2 (T185 + Y187) antibody (ET1610-13) at 1/100 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.

  • Western blot analysis of Phospho-Erk1 (T202 + Y204) + Erk2 (T185 + Y187) on different lysates with Rabbit anti-Phospho-Erk1 (T202 + Y204) + Erk2 (T185 + Y187) antibody (<a href="/products/ET1610-13" style="font-weight: bold;text-decoration: underline;">ET1610-13</a>) at 1/5,000 dilution and competitor's antibody at 1/5,000 dilution.<br /><br />Lane 1: SH-SY5Y cell lysate<br />Lane 2: SH-SY5Y treated with 100ng/mL hβ-NGF for 10 minutes cell lysate<br />Lane 3: PC-12 cell lysate<br />Lane 4: PC-12 treated with 100ng/mL hβ-NGF for 10 minutes cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 41/43 kDa<br />Observed band size: 41/43 kDa<br /><br />Exposure time: 1 minute 50 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/ET1610-13" style="font-weight: bold;text-decoration: underline;">ET1610-13</a>) at 1/5,000 dilution and competitor's antibody at 1/5,000 dilution were used in 5% BSA 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 Phospho-Erk1 (T202 + Y204) + Erk2 (T185 + Y187) on different lysates with Rabbit anti-Phospho-Erk1 (T202 + Y204) + Erk2 (T185 + Y187) antibody (ET1610-13) at 1/5,000 dilution and competitor's antibody at 1/5,000 dilution.

    Lane 1: SH-SY5Y cell lysate
    Lane 2: SH-SY5Y treated with 100ng/mL hβ-NGF for 10 minutes cell lysate
    Lane 3: PC-12 cell lysate
    Lane 4: PC-12 treated with 100ng/mL hβ-NGF for 10 minutes cell lysate

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 41/43 kDa
    Observed band size: 41/43 kDa

    Exposure time: 1 minute 50 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 (ET1610-13) at 1/5,000 dilution and competitor's antibody at 1/5,000 dilution were used in 5% BSA at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

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

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