Overview

  • Product name
  • Description
    Rabbit polyclonal to CD62E
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide surrounding aa 584 of human CD62E

  • Positive control
    • Jurkat cell lysate, mouse small intestine and rat kidney tissue lysate.

Properties

Applications

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.

Application Abreviews Notes
ICC/IF Use at an assay dependent concentration.
IHC-P Use a concentration of 10 - 20 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 1 - 4 µg/ml. Predicted molecular weight: 67 kDa.Can be blocked with CD62E peptide (ab113900).

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.
  • Information by UniProt
  • Database links
  • 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

  • Anti-CD62E antibody (ab18981) + Jurkat cell lysate

    Predicted band size: 67 kDa

  • 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.

  • 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.

  • Anti-CD62E antibody (ab18981) at 2 µg/ml + SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

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

  • ab18981 staining CD62E in Rat brain microvascular endothelial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 1% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (1/50 in 1% BSA) for 18 hours at 4°C. A donkey anti-rabbit DyLight® 650 (IgG; 1/500) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • García Nores GD  et al. CD4+ T cells are activated in regional lymph nodes and migrate to skin to initiate lymphedema. Nat Commun 9:1970 (2018). Read more (PubMed: 29773802) »
  • Jamaly S  et al. Elevated plasma levels of P-selectin glycoprotein ligand-1-positive microvesicles in patients with unprovoked venous thromboembolism. J Thromb Haemost N/A:N/A (2018). Read more (PubMed: 29851269) »
See all 18 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Nov 30 2015

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

Abcam user community

Verified customer

Submitted Nov 19 2013

Question
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

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

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/index.html?datasheet=91361 https://www.abcam.com/index.html?datasheet=91361.

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

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!
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Read More

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

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.

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