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    cd62e-antibody-ab18981.pdf

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Immunology Cell Type Markers CD Adhesion
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Anti-CD62E antibody (ab18981)

  • Datasheet
  • SDS
Reviews (2)Q&A (6)References (26)

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Abpromise

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Find out more.

Western blot - Anti-CD62E antibody (ab18981)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD62E antibody (ab18981)
  • Flow Cytometry - Anti-CD62E antibody (ab18981)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD62E antibody (ab18981)

Key features and details

  • Rabbit polyclonal to CD62E
  • Suitable for: IHC-P, WB, Flow Cyt
  • Reacts with: Mouse, Human
  • Isotype: IgG

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Protein
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Recombinant Human CD62E protein (ab167744)

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Overview

  • Product name

    Anti-CD62E antibody
    See all CD62E primary antibodies
  • Description

    Rabbit polyclonal to CD62E
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Mouse
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human CD62E. Surrounding aa 584.
    Database link: P16581
    (Peptide available as ab113900)

  • Positive control

    • WB: Jurkat cell lysate. Flow Cyt: Jurkat cells. IHC-P: Human placenta and mouse aorta tissues.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    Preservative: 0.03% Proclin 300
    Constituents: PBS, 30% Glycerol (glycerin, glycerine), 0.5% BSA, 0.15% EDTA
  • Concentration information loading...
  • Purity

    Affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Cell Type Markers
    • CD
    • Adhesion
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Leukocyte recruitment
    • Cell adhesion molecules
    • Stem Cells
    • Endothelial Progenitors
    • Endothelial Markers
    • Cardiovascular
    • Cardiovascular Markers
    • Cell Markers
    • Endothelial Cells
    • Cardiovascular
    • Angiogenesis
    • Endothelial Cell Markers

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Immunizing Peptide (Blocking)

    • CD62E peptide (ab113900)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human CD62E protein (ab167744)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab18981 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
IHC-P
Mouse
Human
WB
Human
All applications
Rat
Application Abreviews Notes
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
Use a concentration of 0.5 - 4 µg/ml. Predicted molecular weight: 67 kDa.Can be blocked with CD62E peptide (ab113900).
Flow Cyt
Use at an assay dependent concentration.
Notes
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
Use a concentration of 0.5 - 4 µg/ml. Predicted molecular weight: 67 kDa.Can be blocked with CD62E peptide (ab113900).
Flow Cyt
Use at an assay dependent concentration.

Target

  • Function

    Cell-surface glycoprotein having a role in immunoadhesion. Mediates in the adhesion of blood neutrophils in cytokine-activated endothelium through interaction with PSGL1/SELPLG. May have a role in capillary morphogenesis.
  • Sequence similarities

    Belongs to the selectin/LECAM family.
    Contains 1 C-type lectin domain.
    Contains 1 EGF-like domain.
    Contains 6 Sushi (CCP/SCR) domains.
  • Cellular localization

    Membrane.
  • Target information above from: UniProt accession P16581 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 6401 Human
    • Entrez Gene: 20339 Mouse
    • Entrez Gene: 25544 Rat
    • Omim: 131210 Human
    • SwissProt: P16581 Human
    • SwissProt: Q00690 Mouse
    • SwissProt: P98105 Rat
    • Unigene: 82848 Human
    • Unigene: 5245 Mouse
    • Unigene: 10359 Rat
    see all
  • Alternative names

    • CD26E antibody
    • CD62 antigen-like family member E antibody
    • CD62E antibody
    • E selectin antibody
    • E-selectin antibody
    • ELAM antibody
    • ELAM-1 antibody
    • ELAM1 antibody
    • Endothelial adhesion molecule 1 antibody
    • Endothelial leukocyte adhesion molecule 1 antibody
    • ESEL antibody
    • LECAM2 antibody
    • Leukocyte-endothelial cell adhesion molecule 2 antibody
    • LYAM2_HUMAN antibody
    • Sele antibody
    • Selectin E antibody
    see all

Images

  • Western blot - Anti-CD62E antibody (ab18981)
    Western blot - Anti-CD62E antibody (ab18981)
    Anti-CD62E antibody (ab18981) at 1/1000 dilution + Jurkat cell lysate at 80 µg

    Secondary
    Peroxidase conjugated goat anti-rabbit IgG at 1/10000 dilution

    Predicted band size: 67 kDa
    Observed band size: 67 kDa



    Blocking/Dilution buffer: 5% NFDM (1 hr at RT) / TBST.

    Incubated with the primary antibody overnight at 4°C. Incubated with the secondary antibody for 1 hour at room temperature.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD62E antibody (ab18981)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD62E antibody (ab18981)

    ab18981 IHC staining of human placenta with E-Selectin Polyclonal antibody: Immunohistochemistry of formalin fixed, paraffin-embedded tissue after heat-induced antigen retrieval. 5 μg/ml of antibody used for staining. After incubation with the primary antibody, slides were incubated with biotinylated secondary antibody, followed by alkaline phosphatase-streptavidin and chromogen.

  • Flow Cytometry - Anti-CD62E antibody (ab18981)
    Flow Cytometry - Anti-CD62E antibody (ab18981)

    Flow cytometric analysis of Jurkat cells using ab18981 (red) with negative controls (black and green).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD62E antibody (ab18981)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD62E antibody (ab18981)

    ab18981 IHC staining of mouse aorta from obese diabetic mice with E-Selectin Polyclonal antibody: Immunohistochemistry of paraffin-embedded tissue after heat-induced antigen retrieval. 10-20 μg/mL of antibody used for staining. After incubation with the primary antibody, slides were incubated with biotinylated goat anti-rabbit HRP and visualized with DAB substrate using ABC staining system.

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (26)

    Publishing research using ab18981? Please let us know so that we can cite the reference in this datasheet.

    ab18981 has been referenced in 26 publications.

    • Zhang K  et al. Activation of brain endothelium by Escherichia coli K1 virulence factor cglD promotes polymorphonuclear leukocyte transendothelial migration. Med Microbiol Immunol 208:59-68 (2019). PubMed: 30171337
    • Takeda Y  et al. ROCK2 Regulates Monocyte Migration and Cell to Cell Adhesion in Vascular Endothelial Cells. Int J Mol Sci 20:N/A (2019). PubMed: 30884801
    • Wang M  et al. Role of TLR-4 in anti-ß2-glycoprotein I-induced activation of peritoneal macrophages and vascular endothelial cells in mice. Mol Med Rep 19:4353-4363 (2019). PubMed: 30942412
    • Xu G  et al. Dose-Dependent Influences of Ethanol on Ischemic Stroke: Role of Inflammation. Front Cell Neurosci 13:6 (2019). PubMed: 30853895
    • Wu Y  et al. Silencing of RAD51AP1 suppresses epithelial-mesenchymal transition and metastasis in non-small cell lung cancer. Thorac Cancer 10:1748-1763 (2019). PubMed: 31317661
    View all Publications for this product

    Customer reviews and Q&As

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

    1-8 of 8 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-CD62E antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (293-T)
    Permeabilization
    Yes - .1% triton
    Specification
    293-T
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 30 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-CD62E antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Sample
    Rat Cell (Rat Brain Microvascular Endothelial Cells (RBMVEC))
    Specification
    Rat Brain Microvascular Endothelial Cells (RBMVEC)
    Permeabilization
    No
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 19 2013

    Question

    Please provide the IHC protocol

    Read More

    Abcam community

    Verified customer

    Asked on May 13 2013

    Answer

    1. Deparaffinize/hydrate sections:
    a) Incubate sections in three washes of xylene for 5 minutes each.
    b) Incubate sections in two washes of 100% ethanol for 10 minutes each.
    c) Incubate sections in two washes of 95% ethanol for 10 minutes each.
    2. Wash sections twice in ddH2O for 5 minutes each.
    3. Wash sections in PBS for 5 minutes.
    4. For antigen unmasking, heat sections in 10 mM sodium citrate buffer (pH 6.0) for 1 minute at full power followed by 9 minutes at medium power. (Keep slides fully immersed in buffer and maintain temperature at or just below boiling.) Cool slides for 20 minutes after antigen unmasking.
    5. Wash sections in ddH2O three times for 5 minutes each.
    6. Incubate sections in 1% hydrogen peroxide for 10 minutes.
    7. Wash sections in ddH2O three times for 5 minutes each.
    8. Wash section in PBS for 5 minutes.
    9. Block each section with 100-400 µl blocking solution for 1 hour at room temperature.
    10. Remove blocking solution and add 100-400 µl diluted primary antibody to each section. (Dilute antibody in blocking solution.) Incubate overnight at 4oC.
    11. Remove antibody solution and wash sections in PBS three times for 5 minutes each.
    12. Add 100-400 µl secondary antibody, diluted in blocking solution, to each section. Incubate 30 minutes at room temperature.
    13. If using ABC biotin/avidin method, make ABC reagent according to the manufacturer’s instructions and incubate solution for 30 minutes at room temperature.
    14. Remove secondary antibody solution and wash sections three times with PBS for 5 minutes each.
    15. Add 100-400 µl ABC reagent to each section and incubate for 30 minutes at room temperature.
    16. Remove ABC reagent and wash sections three times in PBS for 5 minutes each.
    17. Add 100-400 µl DAB reagent to each section and monitor staining closely.
    18. As soon as the section turns brown, immerse slides in ddH2O.
    19. If desired, counterstain sections in hematotoxylin for 10 seconds.
    20. Wash sections in ddH2O two times for 5 minutes each.
    21. Dehydrate sections:
    a) Incubate sections in 95% ethanol two times for 10 seconds each.
    b) Repeat in 100% ethanol, incubating sections two times for 10 seconds each.
    c) Repeat in xylene, incubating sections two times for 10 seconds each.
    22. Mount coverslips.

    Read More

    Abcam Scientific Support

    Answered on May 13 2013

    Question

    Dear,
    I ordered the following antibodies and received them at the end of September:
    - Ab54426 (anti-CD62P)
    - Ab91353 (Mouse IgG1 – isotype control)
    - Ab 18981 (anti-CD62E)
    - Ab27478 (rabbit polyclonal IgG)
    - Ab2213 (anti-ICAM1)

    I stored them at +4oC till today, but I have noticed today that your storage instructions recommend to aliquot and store most of the antibodies at -20oC or -80oC.
    Could this have had an effect on my stainings?
    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on Nov 06 2012

    Answer

    Thank you for your enquiry.

    I can confirm that ab18414 can be stored at 4oC as described on the datasheet. This should not be aliquotted and frozen.

    The others I would recommend to aliquot and freeze immediately. Keeping them at 4oC for a few weeks should not significantly affect the staining. However, we would recommend to follow the individual storage instructions provided on the individual datasheets as soon as possible after delivery. This will help to ensure that the antibody remains stable and that we can continue to provide our guarantee.

    I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

    Read More

    Abcam Scientific Support

    Answered on Nov 06 2012

    Question

    Dear,
    We want to proceed and test ab54426 in IHC-P. You can send me the discount code.
    Are these the correct isotype antibodies for the primary antibodies I have chosen:
    - Mouse IgG2a, kappa monoclonal [MG2a-53] - isotype control (Ab18414) for the 1° Ab: Anti-ICAM1 antibody [MEM-111] (Ab2213)
    - Rabbit polyclonal IgG (Ab 27478) for the 1° Ab: Rabbit polyclonal to CD62 E (Ab 18981)
    - Mouse IgG1 [ICIGG1] - Isotype Control (ab91353) for the 1° Ab: Anti-CD62P antibody (Ab54426)
    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on Sep 24 2012

    Answer

    I am very pleased to hear you would like to accept our offer and test ab54426 in IHC-P.

    DISCOUNT CODE: ######

    Expiration date: ######

    This code will give you: 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for ab54426 used in IHC-P and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.

    For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

    Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways:

    1) Call to place your order and mention the code to our customer service department;
    2) Include the code in your fax order;
    3) Place your order on the web and enter the promotional code.

    With regards to the isotype controls:
    For ab2213, the light chain type is unknown, so I would recommend to use a general IgG2a isotype control rather than a specific IgG kappa chain isotype control. For example the following product:

    ab91361 Mouse IgG2a [ICIGG2A] - Isotype Control
    https://www.abcam.com/Mouse-IgG2a-Kappa-Monoclonal-B128-Isotype-Control-ab91361.html https://www.abcam.com/Mouse-IgG2a-Kappa-Monoclonal-B128-Isotype-Control-ab91361.html.

    However, please note this is not tested in IHC-P. Unfortunately, I am sorry we do not have any suitable isotype controls tested in IHC-P in this case.

    ab27478 is a suitable isotype control for ab18981
    ab91353 is a suitable isotype control for ab54426

    Any feedback that you can provide regarding ab54426 will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

    The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

    Read More

    Abcam Scientific Support

    Answered on Sep 24 2012

    Question

    WB with HUVEC cells
    sees band at 35 kDa instead of 67 kDa
    also high background
    Ab: 4 ug/ml
    2nd: usually OK
    block: 5% milk
    not sure if reducing conditions were used
    suggestions: use less 1st and 2nd Abs, add reducing agent

    Read More

    Abcam community

    Verified customer

    Asked on Jul 31 2012

    Answer

    Thank you for contacting us.
    I am sorry to hear that the antibody does not seem to work as expected.

    As we had discussed, I would suggest to try the following tips to reduce the background as well as shift the MW to a higher band:

    1) use less primary and/or secondary antibody
    2) add fresh reducing agent to the loading buffer (e.g. DTT or beta mercaptoethanol)

    Please let me know if these suggestions are of help or not. If not, we can discuss other options to resolve the problem.

    I look forward to hear back from you.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Jul 31 2012

    Question


    Thank you for your quick reply.

    Yes, we have the band in 67kDa, too.

    However we don’t have the negative control because our sample is the whole brain of rats. We sacrificed the rats and took out the brain, then homogenized with lysis buffer. We also perform centrifuge and using the supernatant as a sample.

    That’s why we don’t have the negative control.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 14 2012

    Answer



    I agree with Heather that it would be useful to have a negative control sample. I may be mistaken but if E-selectin is only expressed exclusively in endothelial cells, perhaps using an extract from a different cell type as a negative control will be helpful. Alternatively, it may be helpful to look at the literature where E-selectin is blotted for to see what controls others have used.

    Also I am happy to look at your western blot if you'd like to send it to me as an attachment.

    I hope this is helpful. Please contact me again if you have any further questions.

    Read More

    Abcam Scientific Support

    Answered on Jul 14 2012

    Question

    WB: sees 3 bands peptide + Ab (o/n, 4 degree): no band diluted Ab (as control, o/n, 4 degree): also no band --> probably due to absorption of Ab to tube wall peptide + Ab (30 min, RT): same bands as with Ab only --> probably binding equil. not yet established peptide: 1 ug/ml and Ab: 0.5 ug/ml (2-fold excess) suggestion: use peptide in 10-fold excess, incubate 1-2 h at RT peptide with Ab, don't use Ab-control simultaneously

    Read More

    Abcam community

    Verified customer

    Asked on Nov 16 2011

    Answer

    Thank you for your phone call. I just wanted to follow up with you and send you the suggestions we had discussed over the phone: 1) use peptide in 10-fold excess 2) incubate peptide with Ab for 1-2 h at RT before using for WB 3) avoid preparing an "antibody only"-control simultaneously with the peptide-antibody, just use fresh antibody for doing the WB (due to potential absorption of the antibody to the tube wall which will lead to a lower antibody concentration in this tube). I hope this information is helpful to you. Please let me know how everything works out. I wish you good luck. 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

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