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ERK1/2 Recombinant Rabbit Monoclonal Antibody [SA43-03]

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Specification

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Catalog# ET1601-29

ERK1/2 Recombinant Rabbit Monoclonal Antibody [SA43-03]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IP

  • FC

  • IHC-P

  • IHC-Fr

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Predicted reactivity

  • Zebrafish

Predicted species support after-sales service
Conjugation

  • unconjugated

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

Overview

Product Name

ERK1/2 Recombinant Rabbit Monoclonal Antibody [SA43-03]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human ERK2 aa 180-379.

Species Reactivity

Human, Mouse, Rat (Predicted: Zebrafish)

Predicted species support after-sales service

Validated Applications

WB, IF-Cell, IF-Tissue, IP, FC, IHC-P, IHC-Fr

Molecular Weight

Predicted band size: 43/41 kDa

Positive Control

HeLa cell lysate, Jurkat cell lysate, HepG2 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, C6 cell lysate, Jurkat, human breast carcinoma tissue, mouse esophagus tissue, mouse stomach tissue, Hela, MCF-7, NIH/3T3, A549.

Conjugation

unconjugated

Clone Number

SA43-03

RRID

AB_3069611

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), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000-1:20,000

  • IF-Cell

  • 1:2,000

  • IHC-P

  • 1:20,000

  • IHC-Fr

  • 1:200-1:500

  • IF-Tissue

  • 1:500

  • FC

  • 1:1,000

  • IP

  • Use at an assay dependent concentration.

Target

Function

The first mitogen-activated protein kinase to be discovered was ERK1 (MAPK3) in mammals. Since ERK1 and its close relative ERK2 (MAPK1) are both involved in growth factor signaling, the family was termed "mitogen-activated". With the discovery of other members, even from distant organisms (e.g. plants), it has become increasingly clear that the name is a misnomer, since most MAPKs are actually involved in the response to potentially harmful, abiotic stress stimuli (hyperosmosis, oxidative stress, DNA damage, low osmolarity, infection, etc.). Because plants cannot "flee" from stress, terrestrial plants have the highest number of MAPK genes per organism ever found[citation needed]. Thus the role of mammalian ERK1/2 kinases as regulators of cell proliferation is not a generic, but a highly specialized function.

Background References

1. Ye, Q. et al. 2014. Lactoferrin deficiency promotes colitis-associated colorectal dysplasia in mice. PloS one. 9: e103298.

2. Polidoro, L. et al. 2013. Vitamin D protects human endothelial cells from H O oxidant injury through the Mek/Erk-Sirt1 axis activation. Journal of cardiovascular translational research. 6: 221-31.

Sequence Similarity

Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.

Post-translational Modification

Phosphorylated upon KIT and FLT3 signaling (By similarity). Dually phosphorylated on Thr-202 and Tyr-204, which activates the enzyme. Ligand-activated ALK induces tyrosine phosphorylation. Dephosphorylated by PTPRJ at Tyr-204.

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: P27361 Human

SwissProt: P28482 Human

SwissProt: P63085 Mouse

SwissProt: Q63844 Mouse

SwissProt: P21708 Rat

SwissProt: P63086 Rat

Synonyms

ERK 1 antibody

ERK 2 antibody

ERK-2 antibody

ERK1 antibody

erk1/2 antibody

ERK2 antibody

ERT1 antibody

ERT2 antibody

Extracellular signal regulated kinase 1 antibody

Extracellular signal-regulated kinase 2 antibody

Expand

Images

  • Western blot analysis of ERK1/2 on different lysates with Rabbit anti-ERK1/2 antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: Jurkat cell lysate<br />Lane 3: HepG2 cell lysate<br />Lane 4: NIH/3T3 cell lysate<br />Lane 5: PC-12 cell lysate<br />Lane 6: C6 cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 43/41 kDa<br />Observed band size: 43/41 kDa<br /><br />Exposure time: 19 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/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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 ERK1/2 on different lysates with Rabbit anti-ERK1/2 antibody (ET1601-29) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: Jurkat cell lysate
    Lane 3: HepG2 cell lysate
    Lane 4: NIH/3T3 cell lysate
    Lane 5: PC-12 cell lysate
    Lane 6: C6 cell lysate

    Lysates/proteins at 15 µg/Lane.

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

    Exposure time: 19 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 (ET1601-29) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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 Jurkat cells labeling ERK1/2 with Rabbit anti-ERK1/2 antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>) at 1/2,000 dilution and competitor's antibody at 1/1,600 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-ERK1/2 antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>) at 1/2,000 dilution and competitor's antibody at 1/1,600 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 Jurkat cells labeling ERK1/2 with Rabbit anti-ERK1/2 antibody (ET1601-29) at 1/2,000 dilution and competitor's antibody at 1/1,600 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-ERK1/2 antibody (ET1601-29) at 1/2,000 dilution and competitor's antibody at 1/1,600 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;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of ERK1/2 on different lysates with Rabbit anti-ERK1/2 antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>) at 1/5,000 dilution.<br /><br />Lane 1: HEK-293-si NT cell lysate<br />Lane 2: HEK-293-si ERK1/2 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 41/43 kDa<br />Observed band size: 41/43kDa<br /><br />Exposure time: 20 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>, 1/5,000) and Loading control antibody (Rabbit anti-GAPDH, <a href="/products/ET1601-4" style="font-weight: bold;text-decoration: underline;">ET1601-4</a>, 1/10,000) were 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.

    ☑ Knockdown (KD)

    Western blot analysis of ERK1/2 on different lysates with Rabbit anti-ERK1/2 antibody (ET1601-29) at 1/5,000 dilution.

    Lane 1: HEK-293-si NT cell lysate
    Lane 2: HEK-293-si ERK1/2 cell lysate

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 20 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1601-29, 1/5,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were 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 Jurkat cells labeling ERK1/2.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>, red) at 1/1,000 dilution and competitor's antibody (red) at 1/800 dilution, 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 Jurkat cells labeling ERK1/2.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1601-29, red) at 1/1,000 dilution and competitor's antibody (red) at 1/800 dilution, 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).

  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-ERK1/2 antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>) at 1/20,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-ERK1/2 antibody (ET1601-29) at 1/20,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-29) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse esophagus tissue with Rabbit anti-ERK1/2 antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>) at 1/20,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse esophagus tissue with Rabbit anti-ERK1/2 antibody (ET1601-29) at 1/20,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-29) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-ERK1/2 antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>) at 1/20,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-ERK1/2 antibody (ET1601-29) at 1/20,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-29) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat esophagus tissue with Rabbit anti-ERK1/2 antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat esophagus tissue with Rabbit anti-ERK1/2 antibody (ET1601-29) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-29) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat stomach tissue with Rabbit anti-ERK1/2 antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>) at 1/3,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a>) at 1/3,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat stomach tissue with Rabbit anti-ERK1/2 antibody (ET1601-29) at 1/3,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-29) at 1/3,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Application: IHC-Fr<br /><br />Species: Mouse<br /><br />Site: Hippocampus<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/200<br /><br />Antigen retrieval: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.

    Application: IHC-Fr

    Species: Mouse

    Site: Hippocampus

    Sample: Frozen section

    Antibody concentration: 1/200

    Antigen retrieval: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.

  • Application: IHC-Fr<br /><br />Species: Mouse<br /><br />Site: Cerebral cortex<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/200<br /><br />Antigen retrieval: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.

    Application: IHC-Fr

    Species: Mouse

    Site: Cerebral cortex

    Sample: Frozen section

    Antibody concentration: 1/200

    Antigen retrieval: The section was pre-treated using 1% SDS buffer (in PBS, pH 7.4) for 5 minutes at room temperature.

  • ERK1/2 was immunoprecipitated in 0.2mg Jurkat cell lysate with <a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a> at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (<a href="/products/NBI01H" style="font-weight: bold;text-decoration: underline;">NBI01H</a>) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: Jurkat cell lysate (input)<br />Lane 2: Rabbit IgG instead of <a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a> in Jurkat cell lysate<br />Lane 3: <a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</a> IP in Jurkat cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 43 seconds; ECL: K1801

    ERK1/2 was immunoprecipitated in 0.2mg Jurkat cell lysate with ET1601-29 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1601-29 at 1/5,000 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: Jurkat cell lysate (input)
    Lane 2: Rabbit IgG instead of ET1601-29 in Jurkat cell lysate
    Lane 3: ET1601-29 IP in Jurkat cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 43 seconds; ECL: K1801

  • Immunocytochemistry analysis of PC-12 cells labeling ERK1/2 with Rabbit anti-ERK1/2 antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</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-ERK1/2 antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</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 PC-12 cells labeling ERK1/2 with Rabbit anti-ERK1/2 antibody (ET1601-29) 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-ERK1/2 antibody (ET1601-29) 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.

  • Immunocytochemistry analysis of NIH/3T3 cells labeling ERK1/2 with Rabbit anti-ERK1/2 antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</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-ERK1/2 antibody (<a href="/products/ET1601-29" style="font-weight: bold;text-decoration: underline;">ET1601-29</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 NIH/3T3 cells labeling ERK1/2 with Rabbit anti-ERK1/2 antibody (ET1601-29) 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-ERK1/2 antibody (ET1601-29) 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.

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

Citation

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