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  1. Link

    elk1-antibody-e277-ab32106.pdf

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Signal Transduction Protein Phosphorylation Ser / Thr Kinases MAPK Pathway
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ELK1 antibody [E277] (ab32106)

  • Datasheet
  • SDS
Submit a review Q&A (1)References (39)

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Western blot - Anti-ELK1 antibody [E277] (ab32106)
  • Immunocytochemistry/ Immunofluorescence - Anti-ELK1 antibody [E277] (ab32106)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ELK1 antibody [E277] (ab32106)
  • Flow Cytometry (Intracellular) - Anti-ELK1 antibody [E277] (ab32106)
  • Western blot - Anti-ELK1 antibody [E277] (ab32106)
  • Western blot - Anti-ELK1 antibody [E277] (ab32106)
  • Anti-ELK1 antibody [E277] (ab32106)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E277] to ELK1
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

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Alexa Fluor® 488 Carrier Free

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Overview

  • Product name

    Anti-ELK1 antibody [E277]
    See all ELK1 primary antibodies
  • Description

    Rabbit monoclonal [E277] to ELK1
  • Host species

    Rabbit
  • Specificity

    This antibody does not cross react with other ETS family members
  • Tested applications

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

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Epitope

    ab32106 reacts with an epitope located in the C terminal region of ELK1.
  • Positive control

    • IHC-P: Human breast carcinoma ICC/IF: A549 cells WB: HeLa, MCF7 and HAP1 cell lysates. Flow Cyt (intra): HeLa cells.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    E277
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • MAPK Pathway
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Transcription factors

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-ELK1 antibody [E277] (ab224929)
    • Anti-ELK1 antibody [E277] - BSA and Azide free (ab227114)
  • Assay kits

    • Elk1 Transcription Factor Assay Kit (Colorimetric) (ab207202)
  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human ELK1 knockout HeLa cell line (ab261764)
  • KO cell lysates

    • Human ELK1 knockout HeLa cell lysate (ab256904)
  • Recombinant Protein

    • Recombinant Human ELK1 protein (ab70380)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32106 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
Flow Cyt (Intra)
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
1/500. Detects a band of approximately 62 kDa (predicted molecular weight: 45 kDa).
IHC-P
1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/500.
Notes
Flow Cyt (Intra)
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
1/500. Detects a band of approximately 62 kDa (predicted molecular weight: 45 kDa).
IHC-P
1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/500.

Target

  • Function

    Stimulates transcription. Binds to purine-rich DNA sequences. Can form a ternary complex with the serum response factor and the ETS and SRF motifs of the fos serum response element.
  • Tissue specificity

    Lung and testis.
  • Sequence similarities

    Belongs to the ETS family.
    Contains 1 ETS DNA-binding domain.
  • Post-translational
    modifications

    Sumoylation represses transcriptional activator activity as it results in recruitment of HDAC2 to target gene promoters which leads to decreased histone acetylation and reduced transactivator activity. It also regulates nuclear retention.
    On mitogenic stimulation, phosphorylated on C-terminal serine and threonine residues by MAPK1. Ser-383 and Ser-389 are the preferred sites for MAPK1. In vitro, phosphorylation by MAPK1 potentiates ternary complex formation with the serum responses factors, SRE and SRF. Phosphorylation leads to loss of sumoylation and restores transcriptional activator activity.
  • Cellular localization

    Nucleus.
  • Target information above from: UniProt accession P19419 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2002 Human
    • Entrez Gene: 13712 Mouse
    • Entrez Gene: 314436 Rat
    • Omim: 311040 Human
    • SwissProt: P19419 Human
    • SwissProt: P41969 Mouse
    • SwissProt: A4GTP4 Rat
    • Unigene: 181128 Human
    • Unigene: 405823 Mouse
    see all
  • Alternative names

    • ELK 1 antibody
    • Elk1 antibody
    • ELK1 member of ETS oncogene family antibody
    • ELK1 protein antibody
    • ELK1, ETS transcription factor antibody
    • ELK1_HUMAN antibody
    • ELK2 member of ETS oncogene family antibody
    • ETS domain containing protein Elk 1 antibody
    • ETS domain containing protein Elk1 antibody
    • ETS domain protein Elk1 antibody
    • ETS domain-containing protein Elk-1 antibody
    • ETS like gene 1 antibody
    • Member of ETS oncogene family antibody
    • Oncogene Elk1 antibody
    • Tyrosine kinase (ELK1) oncogene antibody
    see all

Images

  • Western blot - Anti-ELK1 antibody [E277] (ab32106)
    Western blot - Anti-ELK1 antibody [E277] (ab32106)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: ELK1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa positive whole cell lysate (21 µg)
    Lane 4: MCF7 negative whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab32106 observed at 55 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab32106 was shown to specifically react with ELK1 in wild-type HAP1 cells as signal was lost in ELK1 knockout cells. Wild-type and ELK1 knockout samples were subjected to SDS-PAGE. Ab32106 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-ELK1 antibody [E277] (ab32106)
    Immunocytochemistry/ Immunofluorescence - Anti-ELK1 antibody [E277] (ab32106)

    Immunocytochemistry/ Immunofluorescence analysis of A549 (human lung carcinoma epithelial cell) cells labeling ELK1 with purified ab32106 at 1/500 dilution (3.8 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ELK1 antibody [E277] (ab32106)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ELK1 antibody [E277] (ab32106)

    Immunohistochemical analysis of paraffin embedded human breast carcinoma using ab32106 at a dilution of 1/50

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Flow Cytometry (Intracellular) - Anti-ELK1 antibody [E277] (ab32106)
    Flow Cytometry (Intracellular) - Anti-ELK1 antibody [E277] (ab32106)

    Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling ELK1 with unpurified ab32106 at 1/20 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. 

     

     

  • Western blot - Anti-ELK1 antibody [E277] (ab32106)
    Western blot - Anti-ELK1 antibody [E277] (ab32106)
    All lanes : Anti-ELK1 antibody [E277] (ab32106) at 1/500 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ELK1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 45 kDa
    Observed band size: 55 kDa why is the actual band size different from the predicted?



    Lanes 1- 2: Merged signal (red and green). Green - ab32106 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab32106 was shown to react with ELK1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261764 (knockout cell lysate ab256904) was used. Wild-type HeLa and ELK1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32106 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-ELK1 antibody [E277] (ab32106)
    Western blot - Anti-ELK1 antibody [E277] (ab32106)
    Anti-ELK1 antibody [E277] (ab32106) at 1/500 dilution
    Predicted band size: 45 kDa
    Observed band size: 62 kDa why is the actual band size different from the predicted?

  • Anti-ELK1 antibody [E277] (ab32106)
    Anti-ELK1 antibody [E277] (ab32106)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (39)

Publishing research using ab32106? Please let us know so that we can cite the reference in this datasheet.

ab32106 has been referenced in 39 publications.

  • Zhang Q  et al. Transcription factor ELK1 accelerates aerobic glycolysis to enhance osteosarcoma chemoresistance through miR-134/PTBP1 signaling cascade. Aging (Albany NY) 13:6804-6819 (2021). PubMed: 33621196
  • Weber H  et al. MED19 alters AR occupancy and gene expression in prostate cancer cells, driving MAOA expression and growth under low androgen. PLoS Genet 17:e1008540 (2021). PubMed: 33513133
  • Ma J  et al. c-KIT-ERK1/2 signaling activated ELK1 and upregulated carcinoembryonic antigen expression to promote colorectal cancer progression. Cancer Sci 112:655-667 (2021). PubMed: 33247506
  • Ribas VT  et al. AAV-mediated inhibition of ULK1 promotes axonal regeneration in the central nervous system in vitro and in vivo. Cell Death Dis 12:213 (2021). PubMed: 33637688
  • Ni S  et al. EZH2 Mediates miR-146a-5p/HIF-1a to Alleviate Inflammation and Glycolysis after Acute Spinal Cord Injury. Mediators Inflamm 2021:5591582 (2021). PubMed: 34104112
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

Question

The lot number of ELK-1 antibody is YJ060812DS.   

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Abcam community

Verified customer

Asked on Jun 10 2013

Answer

The concentration of this lot is 0.2180 mg/ml

Read More

Abcam Scientific Support

Answered on Jun 10 2013

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