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Anti-EEA1 antibody (ab50313)

Reviews (2)Q&A (4)References (18)
Immunocytochemistry/ Immunofluorescence - Anti-EEA1 antibody (ab50313)
  • Immunocytochemistry/ Immunofluorescence - Anti-EEA1 antibody (ab50313)

Key features and details

  • Rabbit polyclonal to EEA1
  • Suitable for: ICC/IF
  • Reacts with: Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (ab109110)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-EEA1 antibody
    See all EEA1 primary antibodies

  • Description

    Rabbit polyclonal to EEA1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide:

    GSQGSDLDSSATPINTVDVNNESSSEG

    conjugated to KLH by a C-terminal Cysteine residue linker, corresponding to amino acids 24 - 40 of Human EEA1
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab50313 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 (1)
Use a concentration of 5 - 10 µg/ml.
Notes
ICC/IF
Use a concentration of 5 - 10 µg/ml.

Target

  • Function

    Binds phospholipid vesicles containing phosphatidylinositol 3-phosphate and participates in endosomal trafficking.

  • Sequence similarities

    Contains 1 C2H2-type zinc finger.
    Contains 1 FYVE-type zinc finger.

  • Domain

    The FYVE-type zinc finger domain mediates interactions with phosphatidylinositol 3-phosphate in membranes of early endosomes and penetrates bilayers. The FYVE domain insertion into PtdIns(3)P-enriched membranes is substantially increased in acidic conditions.

  • Cellular localization

    Cytoplasm. Early endosome membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • Early endosome antigen 1 antibody
      • Early endosome antigen 1, 162kD antibody
      • Early endosome associated protein antibody
      • EEA 1 antibody
      • EEA1 antibody
      • EEA1_HUMAN antibody
      • Endosome associated protein p162 antibody
      • Endosome-associated protein p162 antibody
      • MST105 antibody
      • MSTP105 antibody
      • ZFYVE2 antibody
      • Zinc finger FYVE domain containing protein 2 antibody
      • Zinc finger FYVE domain-containing protein 2 antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-EEA1 antibody (ab50313)
      Immunocytochemistry/ Immunofluorescence - Anti-EEA1 antibody (ab50313)
      HeLa cells fixed and permeabilized with methanol followed by acetone. Fixed cells were stained with 1 µg/mL ab50313."
    • Immunocytochemistry/ Immunofluorescence - Anti-EEA1 antibody (ab50313)
      Immunocytochemistry/ Immunofluorescence - Anti-EEA1 antibody (ab50313)This image is a courtesy of Anonymous Abreview

      ab50313 staining EEA1 in mouse primary astrocytes by Immunocytochemistry. Cells were fixed with paraformaldehyde, permeabilized with 0.05% saponin and blocking with 10% serum was performed for 20 minutes at 250C. Samples were incubated with primary antibody (1/150: in 10% goat serum / PBS) for 1 hour at 25°C. An Abcam`s ab6939, Cy3®-conjugated goat polyclonal to rabbit IgG (H&L) was used as secondary antibody at 1/200 dilution.

      See Abreview

    References (18)

    Publishing research using ab50313? Please let us know so that we can cite the reference in this datasheet.

    ab50313 has been referenced in 18 publications.

    • Tang FL  et al. Coupling of terminal differentiation deficit with neurodegenerative pathology in Vps35-deficient pyramidal neurons. Cell Death Differ 27:2099-2116 (2020). PubMed: 31907392
    • Levanova N  et al. The Legionella effector LtpM is a new type of phosphoinositide-activated glucosyltransferase. J Biol Chem 294:2862-2879 (2019). PubMed: 30573678
    • Martini F  et al. The ESCRT-0 Protein HRS Interacts with the Human T Cell Leukemia Virus Type 2 Antisense Protein APH-2 and Suppresses Viral Replication. J Virol 94:N/A (2019). PubMed: 31597781
    • Leiva-Rodríguez T  et al. ATG5 overexpression is neuroprotective and attenuates cytoskeletal and vesicle-trafficking alterations in axotomized motoneurons. Cell Death Dis 9:626 (2018). PubMed: 29799519
    • Fischer AW  et al. The adaptor protein PID1 regulates receptor-dependent endocytosis of postprandial triglyceride-rich lipoproteins. Mol Metab 16:88-99 (2018). PubMed: 30100244
    View all Publications for this product

    Customer reviews and Q&As

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

    1-6 of 6 Abreviews or Q&A

    Immunocytochemistry abreview for Anti-EEA1 antibody

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Mouse Cultured Cells (primary astrocyte)
    Specification
    primary astrocyte
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.05% saponin
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 08 2009

    Immunocytochemistry/ Immunofluorescence abreview for Anti-EEA1 antibody

     Average
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Lung carcinoma cell line)
    Specification
    Lung carcinoma cell line
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - Triton X-100
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 17 2009

    Question

    Thanks for your email. Please find my answers inserted below.

    I will be happy to send the replacement ab95195, but I will need the original order number or PO number that ab2074 was received on. Would you be able to provide this information? I've found an order from 2007 going to ***; could this be your order? If so, did the antibody work when it was first received? At what temperature has it been stored?

    Yes, it is correct that this is still about our order in 2007. As we discussed last time on the phone, the antibody did not work from the very beginning. Originally, I could not be sure that the antibody was the problem because I did not have transfected cells or control antibody available for a rigorous comparison. We know now that we should have been able to detect endogenous CXCR4 in Hela cells under the conditions used, and we did not in 2007. Therefore, you graciously offered to still send a replacement after this long time. We aliquot our antibodies and store them in two ways: We keep a small aliquot at 4C for immediate testing, another aliquot receives 50% glycerol/PBS and is stored at -20C; the latter seems the best for most antibodies and we prefer it over freezing of multiple small aliquots. Neither aliquot worked. However, I did notice that there was some precipitate at the bottom of the vial when we received it.So, I cannot rule out that is was just the specific batch of CXCR4 antibody you sent that was flawed.

    Thank you also for your feedback regarding the conditions of ab24170. Would you be able to submit an Abreview showing your results under these conditions, to help highlight the optimal conditions? You will receive Abpoints in exchange for submitting the review, and these points can be redeemed for a discount on a future purchase. I've also added your feedback to our notes on this antibody.

    Will do as soon as I find the time.

    I am very sorry to hear that the EEA1 antibody ab50313 did not give satisfactory results, and I can send a replacement of your choice. We have a rabbit monoclonal antibody to this target, ab109110, which is suitable for use with mouse and human samples in ICC/IF:

    https://www.abcam.com/EEA1-antibody-EPR4245-ab109110.html

    Would this antibody work for you? Are there any other targets that you're interested in besides EEA1?

    The data sheet states that ab109110 works for ICC, but there is no image. I would like to hold off with a replacement until I see an image that shows the expected pattern in cultured cells. Also, we spent an enormous amount of time on testing antibodies and had to move on with the project. Thus, we may be more interested in a different epitome in due course.

    Thank you again for your response and I look forward to hearing from you. Please let me know if you have any questions or need anything else at this time.

    Read More
    Answer

    Thank you very much for your responses.
    I'm sending a free of charge vial of ab95195 on the order ***, which should arrive tomorrow. Please keep me updated about the results using this antibody.
    For the replacement of ab50313, I certainly understand that you have spent a lot of time on this target. So, please let me know once you have chosen a replacement and I'll be happy to send that out to you promptly.
    Please let me know if you have any questions or if there is anything else that we can do for you.
    Have a nice day!

    Read More

    Question

    I promised to get back in touch with you several months ago, my apologies that it has taken me this long. Here is some more informationabout three of your antibodies and my request for a second replacement.

    1. I have previously been in contact regarding ab2074, which did not yield specific staining for us under the following conditionsat aconcentration of up to 100 ug/ml (1:200 dilution),while a different antibody (UMB2) gave bright staining:

    - cells transfected with human or rat CXCR4 cDNA, PFA-fixed and permeabilized with TX100
    - Hela cellsPFA-fixed and permeabilized with TX100 or methanol fixed

    You offered a replacement, but I did not yet specify the antibody I would like to receive. I now would like to request anti-APPL antibodyab95195 as a replacement.



    2. Information for anti-lamp-1 antibody ab24170: This antibody works well for us in cells of human or rat origin at 5 ug/ml when using PFA fixation followed by saponin permeabilization. This antibody does NOT work in our hands using TX100 or methanol permeabilization. This finding seems to be typical for antibodies directed to the short cytoplasmic tails of lysosomal membrane proteins (we saw it before with two different antibodies of this kind). Note that another customer used Tween and achieved a labeling pattern consistent with that of lysosomes. It would be helpful to include a note in the data sheet that permeabilizations other than with TX100 or methanol (such as saponin) should be tried if standard procedures are not successful (most frequently TX100 or methanol are used) to avoid frustrations or wrong results with this fine antibody.



    3. Information for anti-EEA1 antibody ab 50313 and request for replacement (to be specified). We received this antibody as a replacement for ab2900 on 8/30/11. This antibody was also not satisfactory for us, although it gave more specific staining (together with strong nonspecific staining). We found that the reactivity of EEA1 antibodies is much better after short fixations (5 min, 4% PFA) than after longer fixations (>30 min and/or higher temperatures) indicating that some epitopes are sensitive to (over)fixation. While we can obtain some EEA1-specific labelingwith ab50313, as evidenced by coincidence staining with a monoclonal ab to EEA1, there are also many structures in our cells that are similarly bright labeled withab50313 that are negative for mAb staining. Therefore, this antibody is also of no use to as we cannot use it with confidence to identify early endosomes. We have been using Flp-In™-293 cells, a subclone of the commonly used human 293 cells.


    Also, please note my new email address.


    Best regards,

    Read More
    Answer

    Thank you very much for your reply and for sending all of this information.
    I will be happy to send the replacement ab95195, but I will need the original order number or PO number that ab2074 was received on. Would you be able to provide this information? I've found an order from 2007; could this be your order? If so, did the antibody work when it was first received? At what temperature has it been stored?
    Thank you also for your feedback regarding the conditions of ab24170. Would you be able to submit an Abreview showing your results under these conditions, to help highlight the optimal conditions? You will receive Abpoints in exchange for submitting the review, and these points can be redeemed for a discount on a future purchase. I've also added your feedback to our notes on this antibody.
    I am very sorry to hear that the EEA1 antibody ab50313 did not give satisfactory results, and I can send a replacement of your choice. We have a rabbit monoclonal antibody to this target, ab109110, which is suitable for use with mouse and human samples in ICC/IF:
    https://www.abcam.com/EEA1-antibody-EPR4245-ab109110.html
    Would this antibody work for you? Are there any other targets that you're interested in besides EEA1?
    Thank you again for your response and I look forward to hearing from you. Please let me know if you have any questions or need anything else at this time.

    Read More

    Question

    My technician is currently doing additional testing with ab50313. Because I will be out of the country until Nov 17, I would prefer to get back to you after my return in conjunction with those new results. I hope this delay is OK, but please let me know of there is a problem with this.

    Read More
    Answer

    Thanks for the update. We certainly appreciate your lab's troubleshooting efforts. Please feel free to reply after your trip.

    Read More

    Question

    High background staining human and rat cells.

    Read More
    Answer

    We have previously been in contact regarding ab2074, which was not providing satisfactory results. We had offered a free of charge replacement with a different antibody, but we were waiting to hear from you with your selection. If you have determined which antibody you would like as a replacement, please let us know and we will send it promptly. I wish you the best of luck with your research and please do not hesitate to contact us if you have any further questions.

    Read More

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