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    products/primary-antibodies/midkine-antibody-ep1143y-ab52637.pdf

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Neuroscience Neurology process Growth and Development Neurotrophins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Midkine antibody [EP1143Y] (ab52637)

  • Datasheet
  • SDS
Reviews (1)Q&A (1)References (16)

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Western blot - Anti-Midkine antibody [EP1143Y] (ab52637)
  • Western blot - Anti-Midkine antibody [EP1143Y] (ab52637)
  • Immunocytochemistry/ Immunofluorescence - Anti-Midkine antibody [EP1143Y] (ab52637)
  • Immunoprecipitation - Anti-Midkine antibody [EP1143Y] (ab52637)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Midkine antibody [EP1143Y] (ab52637)
  • Western blot - Anti-Midkine antibody [EP1143Y] (ab52637)
  • Western blot - Anti-Midkine antibody [EP1143Y] (ab52637)
  • Western blot - Anti-Midkine antibody [EP1143Y] (ab52637)
  • Flow Cytometry (Intracellular) - Anti-Midkine antibody [EP1143Y] (ab52637)
  • Immunocytochemistry/ Immunofluorescence - Anti-Midkine antibody [EP1143Y] (ab52637)
  • Anti-Midkine antibody [EP1143Y] (ab52637)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1143Y] to Midkine
  • Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human, Recombinant fragment

Conjugates logo Related conjugates and formulations

APC Carrier Free HRP PE

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Recombinant mouse Midkine protein (ab123474)
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Isotype
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Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free) (ab199376)

View more associated products

Overview

  • Product name

    Anti-Midkine antibody [EP1143Y]
    See all Midkine primary antibodies
  • Description

    Rabbit monoclonal [EP1143Y] to Midkine
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human, Recombinant fragment
  • Immunogen

    Synthetic peptide corresponding to Human Midkine aa 100 to the C-terminus (C terminal).

  • Positive control

    • Human liver carcinoma; Human pancreatic carcinoma; HeLa cells; SH-SY5Y (Human neuroblastoma cell line from bone marrow).
  • 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.

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

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. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP1143Y
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Growth and Development
    • Neurotrophins
    • Neuroscience
    • Neurology process
    • Neurogenesis

Associated products

  • Alternative Versions

    • Anti-Midkine antibody [EP1143Y] - BSA and Azide free (ab236781)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant mouse Midkine protein (ab123474)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab52637 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)
Use at an assay dependent concentration.
WB (1)
1/1000. Detects a band of approximately 16 kDa (predicted molecular weight: 16 kDa).

For unpurified use at 1/500.

IP
1/20.

For unpurified use at 1/30. 

IHC-P
1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100.

For unpurified use at 1/250 - 1/500.

Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
1/1000. Detects a band of approximately 16 kDa (predicted molecular weight: 16 kDa).

For unpurified use at 1/500.

IP
1/20.

For unpurified use at 1/30. 

IHC-P
1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100.

For unpurified use at 1/250 - 1/500.

Target

  • Function

    Has heparin binding activity, and growth promoting activity. Involved in neointima formation after arterial injury, possibly by mediating leukocyte recruitment. Also involved in early fetal adrenal gland development.
  • Sequence similarities

    Belongs to the pleiotrophin family.
  • Cellular localization

    Secreted.
  • Target information above from: UniProt accession P21741 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4192 Human
    • Omim: 162096 Human
    • SwissProt: P21741 Human
    • Unigene: 82045 Human
    • Alternative names

      • Amphiregulin associated protein antibody
      • Amphiregulin-associated protein antibody
      • ARAP antibody
      • FLJ27379 antibody
      • Mdk antibody
      • Midgestation and kidney protein antibody
      • Midkine antibody
      • MK 1 antibody
      • MK antibody
      • MK_HUMAN antibody
      • MK1 antibody
      • NEGF 2 antibody
      • NEGF2 antibody
      • Neurite growth promoting factor 2 antibody
      • Neurite outgrowth promoting protein antibody
      • Neurite outgrowth-promoting factor 2 antibody
      • Neurite outgrowth-promoting protein antibody
      • Retinoic acid inducible factor antibody
      see all

    Images

    • Western blot - Anti-Midkine antibody [EP1143Y] (ab52637)
      Western blot - Anti-Midkine antibody [EP1143Y] (ab52637)

      ab52637 was shown to react with MDK in wild-type HAP1 cells in Western blot with loss of signal observed in a MDK knockout cell line. Wild-type HAP1 and MDK knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab52637 overnight at 4 °C at a 1/500 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2 μg/ml before imaging. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

    • Western blot - Anti-Midkine antibody [EP1143Y] (ab52637)
      Western blot - Anti-Midkine antibody [EP1143Y] (ab52637)
      Anti-Midkine antibody [EP1143Y] (ab52637) at 1/500 dilution (unpurified) + Recombinant protein at 0.001 µg

      Secondary
      Goat anti-rabbit HRP labeled at 1/2000 dilution

      Predicted band size: 16 kDa
      Observed band size: 16 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-Midkine antibody [EP1143Y] (ab52637)
      Immunocytochemistry/ Immunofluorescence - Anti-Midkine antibody [EP1143Y] (ab52637)

      Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Midkine with purified ab52637 at 1:100 dilution (10.5μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Immunoprecipitation - Anti-Midkine antibody [EP1143Y] (ab52637)
      Immunoprecipitation - Anti-Midkine antibody [EP1143Y] (ab52637)

      ab52637 (purified) at 1:20 dilution (2ug) immunoprecipitating Midkine in SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate.

      Lane 1 (input): SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate 10ug

      Lane 2 (+): ab52637 & SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52637 in SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate

      For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:5000 dilution.

      Blocking and diluting buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Midkine antibody [EP1143Y] (ab52637)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Midkine antibody [EP1143Y] (ab52637)

      Human liver carcinoma stained with unpurified ab52637 at 1/100 - 1/250 dilution

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

    • Western blot - Anti-Midkine antibody [EP1143Y] (ab52637)
      Western blot - Anti-Midkine antibody [EP1143Y] (ab52637)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human pancreatic carcinoma tissue sections labeling Midkine with purified ab52637 at 1:50 dilution (2.1 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ab97051 Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1:500 dilution. PBS instead of the primary antibody was used as the negative control.

    • Western blot - Anti-Midkine antibody [EP1143Y] (ab52637)
      Western blot - Anti-Midkine antibody [EP1143Y] (ab52637)
      Anti-Midkine antibody [EP1143Y] (ab52637) at 1/1000 dilution (purified) + SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 16 kDa
      Observed band size: 16 kDa



      Blocking and diluting: 5% NFDM/TBST

    • Western blot - Anti-Midkine antibody [EP1143Y] (ab52637)
      Western blot - Anti-Midkine antibody [EP1143Y] (ab52637)
      Anti-Midkine antibody [EP1143Y] (ab52637) at 1/10000 dilution (purified) + Recombinant Human Midkine protein (ab49848) at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 16 kDa
      Observed band size: 16 kDa



      Blocking and diluting buffer: 5% NFDM/TBST

    • Flow Cytometry (Intracellular) - Anti-Midkine antibody [EP1143Y] (ab52637)
      Flow Cytometry (Intracellular) - Anti-Midkine antibody [EP1143Y] (ab52637)

      Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Midkine with purified ab52637 at 1/250 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (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. 

       

       

    • Immunocytochemistry/ Immunofluorescence - Anti-Midkine antibody [EP1143Y] (ab52637)
      Immunocytochemistry/ Immunofluorescence - Anti-Midkine antibody [EP1143Y] (ab52637)

      HeLa cells stained with unpurified ab52637 at 1/250 - 1/500 dilution

    • Anti-Midkine antibody [EP1143Y] (ab52637)
      Anti-Midkine antibody [EP1143Y] (ab52637)

    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 (16)

    Publishing research using ab52637? Please let us know so that we can cite the reference in this datasheet.

    ab52637 has been referenced in 16 publications.

    • Johnson ECB  et al. Large-scale deep multi-layer analysis of Alzheimer's disease brain reveals strong proteomic disease-related changes not observed at the RNA level. Nat Neurosci 25:213-225 (2022). IHC-P ; Human . PubMed: 35115731
    • Wu D  et al. Long non-coding RNA maternally expressed gene 3 affects cell proliferation, apoptosis and migration by targeting the microRNA-9-5p/midkine axis and activating the phosphoinositide-dependent kinase/AKT pathway in hepatocellular carcinoma. Oncol Lett 21:345 (2021). PubMed: 33747202
    • Duan R  et al. The Long Noncoding RNA ZFAS1 Potentiates the Development of Hepatocellular Carcinoma via the microRNA-624/MDK/ERK/JNK/P38 Signaling Pathway. Onco Targets Ther 13:4431-4444 (2020). PubMed: 32547074
    • Higginbotham L  et al. Integrated proteomics reveals brain-based cerebrospinal fluid biomarkers in asymptomatic and symptomatic Alzheimer's disease. Sci Adv 6:N/A (2020). PubMed: 33087358
    • Weckbach LT  et al. Midkine drives cardiac inflammation by promoting neutrophil trafficking and NETosis in myocarditis. J Exp Med 216:350-368 (2019). PubMed: 30647120
    View all Publications for this product

    Customer reviews and Q&As

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

    1-2 of 2 Abreviews or Q&A

    Western blot abreview for Anti-Midkine antibody [EP1143Y]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Chicken Tissue lysate - whole (E1.5 embryos)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris)
    Loading amount
    10 µg
    Specification
    E1.5 embryos
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jan 23 2017

    Question

    To who it might concern, Do know the antibody concentration for product cat.number ab52637 (anti-Midkine clone EP1134Y). I cant find the information on your product page and I need it for an in vitro neutralization experiment I about to set up.    

    Read More

    Abcam community

    Verified customer

    Asked on Nov 16 2011

    Answer

    Thank you for contacting us. This product is a tissue culture supernatant, not a purified antibody. Concentrations are generally not determined for tissue culture supernatant products as they are not relevance; they may contain other proteins aside from the antibody of interest. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Nov 16 2011

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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