Overview

  • Product name

  • Description

    Rabbit polyclonal to ESAM
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Cynomolgus monkey
  • Immunogen

    Synthetic peptide corresponding to Human ESAM aa 100-200 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab101730)

  • Positive control

    • This antibody gave a positive signal in the following lysates: Human Heart tissue; Human Liver tissue; Human Kidney tissue; Human Lung tissue; HepG2 whole cell; HEK293 whole cell.

Properties

Applications

Our Abpromise guarantee covers the use of ab74777 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 a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 41 kDa).

Target

  • Function

    Can mediate aggregation most likely through a homophilic molecular interaction.
  • Tissue specificity

    Highly expressed in endothelial cells.
  • Sequence similarities

    Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Cellular localization

    Cell junction > adherens junction. Cell junction > tight junction. Cell membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • 2310008D05Rik antibody
    • Endothelial cell adhesion molecule antibody
    • Endothelial cell selective adhesion molecule antibody
    • Endothelial cell-selective adhesion molecule antibody
    • Esam antibody
    • ESAM_HUMAN antibody
    • HUEL (C4orf1) interacting protein antibody
    • LP4791 protein antibody
    • W117m antibody
    see all

Images

  • All lanes : Anti-ESAM antibody (ab74777) at 1 µg/ml

    Lane 1 : Human heart tissue lysate - total protein (ab29431)
    Lane 2 : Human liver tissue lysate - total protein (ab29889)
    Lane 3 : Human kidney tissue lysate - total protein (ab30203)
    Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 6 : Lung (Human) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Annexin-2/ANXA2 overexpression lysate (ab94080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 41 kDa
    Observed band size: 37,41 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 100 kDa, 15 kDa, 250 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 3 minutes


    The ESAM protein has a predicted molecular weight of 41 kDa. The first 29 amino acids of the ESAM sequence act as a signal sequence. The band at 37 kDa may represent the cleaved form of the ESAM protein.
  • ICC/IF image of ab74777 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab74777, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HepG2 cells at 5µg/ml, and in 100% methanol fixed (5 min) HepG2 and MCF7 cells at 5µg/ml.

References

ab74777 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Sorry I was not able to take your email last week. I was not in the office. I am very sorry that the two anti-ESAM antibodies did not show the staining we had hoped for, it may be that as you say that the epitope presented by the protein (even though these two antibodies are raised against different peptide immunogens) is very difficult to unmask successfully. Thank you for taking the time to try to optimise the staining conditions. It is very much appreciated and will hopefully allow future customers to assess more accurately the applications appropriate to use with these antibodies. I am sorry that the image of the first Abreview you submitted was not published on the website. This was not intentional, it appears that we did not receive the image. This may have been if it was particularly large. If you would be able to send the images for the staining for both ab73988 and ab74777 I can upload these onto the website myself. This will also mean that you get the extra Abpoints for providing images. I look forward to receiving your reply.

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Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Colon tumour)
Specification
Colon tumour
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH6.0
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 08 2011

Answer

Thank you for contacting us. We currently have two antibodies directed towards human EMSA, ab74777 and ab73988. Both are rabbit polyclonals. ab74777 like the antibody you have previously used is directed against the extracellular domain of ESAM while the ab73988 immunogen is taken from the C-terminal domain. Both of the antibodies have been tested by WB, and ICC and IHC frozen sections respectively, however neither of these antibodies have to date been tested for IHC with paraffin sections. If you were to purchase either of these antibodies and use them with human paraffin sections for IHC we would be able to offer you a testing discount. This would mean if you purchase the antibody, test it and share the results with us through an Abreview you would then get a free primary antibody of your choice from our catalogue. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab74777 or ab73988 with IHC-P. I will then send a discount code. This code must be issued before purchasing the antibody so please wait for my reply before ordering. 2. Purchase ab74777 or ab73988 either by phone, fax, or online (www.abcam.com). 3. Test it in IHC-P. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out more about our Abreview system, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab74777 or ab73988 turns out to be unsuitable for IHC-P, you will still receive the discount on your next purchase after your Abreview has been submitted. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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