Overview

  • Product name

    Anti-ERK1 antibody [Y72] - BSA and Azide free
    See all ERK1 primary antibodies
  • Description

    Rabbit monoclonal [Y72] to ERK1 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, Flow Cyt, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human, Recombinant fragment
  • Immunogen

    Synthetic peptide within Human ERK1 aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: P27361
    (Peptide available as ab204281)

  • Epitope

    ab214168 reacts with an epitope located in the N terminal region of ERK1.
  • Positive control

    • WB: HeLa, HEK293, Jurkat and RAW264.7 whole cell lysates and ERK1 recombinant protein. IHC: Human lung carcinoma, human cervix carcinoma and human tonsil tissues. ICC/IF: Jurkat cells. Flow Cyt: HeLa and Jurkat cells. IP: Jurkat cells.
  • General notes

    ab214168 is the carrier-free version of ab32537 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab214168 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab214168 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use at an assay dependent concentration. Detects a band of approximately 44 kDa (predicted molecular weight: 43 kDa).

Can be blocked with ab204281.

IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control.

IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF Use at an assay dependent concentration.

Target

  • Function

    Involved in both the initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors such as ELK-1. Phosphorylates EIF4EBP1; required for initiation of translation. Phosphorylates microtubule-associated protein 2 (MAP2). Phosphorylates SPZ1 (By similarity). Phosphorylates heat shock factor protein 4 (HSF4).
  • Sequence similarities

    Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
    Contains 1 protein kinase domain.
  • Domain

    The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
  • Post-translational
    modifications

    Dually phosphorylated on Thr-202 and Tyr-204, which activates the enzyme. Dephosphorylated by PTPRJ at Tyr-204.
  • Information by UniProt
  • Database links

  • Alternative names

    • ERK 1 antibody
    • ERK-1 antibody
    • ERK1 antibody
    • ERT 2 antibody
    • ERT2 antibody
    • Extracellular Signal Regulated Kinase 1 antibody
    • Extracellular signal related kinase 1 antibody
    • Extracellular signal-regulated kinase 1 antibody
    • HGNC6877 antibody
    • HS44KDAP antibody
    • HUMKER1A antibody
    • Insulin Stimulated MAP2 Kinase antibody
    • Insulin-stimulated MAP2 kinase antibody
    • MAP kinase 1 antibody
    • MAP kinase 3 antibody
    • MAP Kinase antibody
    • MAP kinase isoform p44 antibody
    • MAPK 1 antibody
    • MAPK 3 antibody
    • MAPK antibody
    • MAPK1 antibody
    • Mapk3 antibody
    • MGC20180 antibody
    • Microtubule Associated Protein 2 Kinase antibody
    • Microtubule-associated protein 2 kinase antibody
    • Mitogen Activated Protein Kinase 3 antibody
    • Mitogen-activated protein kinase 1 antibody
    • Mitogen-activated protein kinase 3 antibody
    • MK03_HUMAN antibody
    • OTTHUMP00000174538 antibody
    • OTTHUMP00000174541 antibody
    • p44 ERK1 antibody
    • p44 MAPK antibody
    • p44-ERK1 antibody
    • p44-MAPK antibody
    • P44ERK1 antibody
    • P44MAPK antibody
    • PRKM 3 antibody
    • PRKM3 antibody
    • Protein Kinase Mitogen Activated 3 antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labelling ERK1 with purified ab32537 at a dilution of 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32537).

  • Overlay histogram showing HAP1 wildtype (green line) and HAP1-MAPK3 knockout cells (red line) stained with ab32537. The cells were fixed with  80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab32537,1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 min at 22°C.
    A Rabbit IgG isotype control antibody  (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-MAPK3  knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
    Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32537).

  • ab32537 staining ERK1 in wild-type HAP1 cells (top panel) and ERK1 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab32537 at 1/400 dilution and ab7291 at 1ug/ml concentration overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150117) at 2ug/ml (shown in pseudo-color red). Nuclear DNA was labelled in blue with DAPI.

    This product also gave a positive signal under the same testing conditions in HAP1 cells fixed with 4% formaldehyde (10 min).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32537).

  • ab32537 (purified) at a dilution of 1/20 immunoprecipitating ERK1 in Jurkat whole cell lysate.

    Lane 1 (input): Jurkat whole cell lysate (10µg)

    Lane 2 (+): ab32537 + Jurkat whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32537 in Jurkat whole cell lysate.

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32537).

  • Flow Cytometry analysis of Jurkat cells labelling ERK1 with purified ab32537 at a dilution of 1/30 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32537).

  • Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling ERK1 with purified ab32537 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32537).

  • Overlay histogram showing HeLa cells stained with ab32537 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween 20 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32537, 1/11312) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730, 0.01μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween 20 for 20 min used under the same conditions.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32537).

  • IHC image of ab32537 staining ERK1 in Human tonsil formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32537, 2μg/ml dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32537).

  • Unpurified ab32537 staining ERK1 (green) in HEK293 cells by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized with 70% methanol. The sample was incubated with the primary antibody (1/40 in PBS + 0.2% BSA + 0.1% sodium azide) for 1 hour at 22°C. A phycoerythrin-conjugated goat anti-rabbit IgG (1/100) was used as the secondary antibody.
    Gating Strategy: Live Cells. Purple plot represents isotype control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32537).

  • Unpurified ab32537 staining ERK1 in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100 in PBS. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. ab150081, a goat anti rabbit Alexa Fluor® 488 (1/200) was used as the secondary antibody. Counterstained with DAPI. Cytoplasmic and nuclear staining shown.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32537).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling ERK1 with unpurified ab32537.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32537).

References

This product has been referenced in:

  • Clift D  et al. A Method for the Acute and Rapid Degradation of Endogenous Proteins. Cell 171:1692-1706.e18 (2017). Read more (PubMed: 29153837) »
  • Tor YS  et al. Induction of Apoptosis in MCF-7 Cells via Oxidative Stress Generation, Mitochondria-Dependent and Caspase-Independent Pathway by Ethyl Acetate Extract of Dillenia suffruticosa and Its Chemical Profile. PLoS One 10:e0127441 (2015). WB ; Human . Read more (PubMed: 26047480) »
See all 3 Publications for this product

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