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Anti-E. coli LPS antibody [2D7/1] (ab35654)

Reviews (8)Q&A (24)References (56)
Immunocytochemistry/ Immunofluorescence - Anti-E. coli LPS antibody [2D7/1] (ab35654)

    Key features and details

    • Mouse monoclonal [2D7/1] to E. coli LPS
    • Suitable for: ICC/IF, ELISA
    • Reacts with: Escherichia coli
    • Isotype: IgG2b

    Overview

    • Product name

      Anti-E. coli LPS antibody [2D7/1]

    • Description

      Mouse monoclonal [2D7/1] to E. coli LPS

    • Host species

      Mouse

    • Specificity

      The J5 mutant of E. coli lacks the enzyme uridine diphosphate glucose 4-epimerase and therefore produces an incomplete LPS, deficient in galactose and all the sugars distal to the central polymers. The J5 mutant has no O-specific chains and its endotoxin remains as the core LPS containing lipid A, N acetyl glucosamine, 2-keto-3-deoxyoctonate, heptose and glucose (a composition similar to that of the Rc strains of Salmonella).

      The clone 2D7/1 has also been found to react with K. pneumoniae, S. sonnei and S. typhimurium.

    • Tested applications

      Suitable for: ICC/IF, ELISAmore details

    • Species reactivity

      Reacts with: Escherichia coli

    • Immunogen

      Tissue, cells or virus corresponding to Escherichia coli E. coli LPS. Escherichia coli J5 cells.

    • 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 ab35654 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 (3)
    1/200.
    ELISA (1)
    1/1500.
    Notes
    ICC/IF
    1/200.
    ELISA
    1/1500.

    Target

    • Relevance

      Some form of LPS is a major component of the cell membrane of Gram negative bacteria, contributing greatly to the structural integrity of the bacteria, and protecting the membrane from certain kinds of chemical attack. LPS is an endotoxin, and induces a strong response from normal animal immune systems. LPS function has been under experimental research for several years due to its role in activating many transcriptional factors, which become active after stimulation with LPS. LPS also produces many types of mediators involved in septic shock.

    • Alternative names

      • Escherichia coli LPS antibody

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-E. coli LPS antibody [2D7/1] (ab35654)
      Immunocytochemistry/ Immunofluorescence - Anti-E. coli LPS antibody [2D7/1] (ab35654)This image is courtesy of an anonymous Abreview

      ab35654 staining E. coli LPS in mouse hepatocytes (red). Cells were PFA fixed and permeabilized in 0.1% Triton X-100 prior to blocking in SFX Kit blocking agent for 30 minutes at 25°C. The primary antibody was diluted 1/200 and incubated with the sample for 1 hour at 25°C. The secondary antibody was Alexa Fluor® 555 goat anti-mouse, diluted 1/100. Cells were stimulated for 90 minutes with E. coli LPS and then probed with ab35654. The picture was taken on a Leica confocal microscope that pseudo-colors the 555 secondary red. The blue staining was with To-Pro-III Iodide,again, pseudo-colored to be blue on confocal.

      See Abreview

    References (56)

    Publishing research using ab35654? Please let us know so that we can cite the reference in this datasheet.

    ab35654 has been referenced in 56 publications.

    • McGrath CJ  et al. Development of a novel human intestinal model to elucidate the effect of anaerobic commensals on Escherichia coli infection. Dis Model Mech 15:N/A (2022). PubMed: 35302159
    • Northrop-Albrecht EJ  et al. Assessment of extracellular vesicle isolation methods from human stool supernatant. J Extracell Vesicles 11:e12208 (2022). PubMed: 35383410
    • Fu A  et al. Tumor-resident intracellular microbiota promotes metastatic colonization in breast cancer. Cell 185:1356-1372.e26 (2022). PubMed: 35395179
    • Ma XY  et al. Citation: Tight Junction Protein Expression-Inducing Probiotics Alleviate TNBS-Induced Cognitive Impairment with Colitis in Mice. Nutrients 14:N/A (2022). PubMed: 35889931
    • Chen X  et al. Intracellular galectin-3 is a lipopolysaccharide sensor that promotes glycolysis through mTORC1 activation. Nat Commun 13:7578 (2022). PubMed: 36481721
    View all Publications for this product

    Customer reviews and Q&As

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

    1-10 of 32 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-E. coli LPS antibody [2D7/1]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Colon (infected with E.coli))
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
    Permeabilization
    No
    Specification
    Colon (infected with E.coli)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Fixative
    10% normal buffered formalin
    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 Dec 11 2018

    Immunocytochemistry/ Immunofluorescence abreview for Anti-E. coli LPS antibody [2D7/1]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Drosophila melanogaster Cell (hemocyte with engulfed E. coli (DH5a))
    Permeabilization
    Yes - 0.1% Triton
    Specification
    hemocyte with engulfed E. coli (DH5a)
    Blocking step
    Serum as blocking agent for 15 minute(s) · Concentration: 5% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    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.

    Catherine Brennan

    Verified customer

    Submitted Jan 10 2018

    Immunocytochemistry/ Immunofluorescence abreview for Anti-E. coli LPS antibody [2D7/1]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Hela)
    Permeabilization
    Yes - TritonX-100
    Specification
    Hela
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    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 May 19 2015

    ELISA abreview for Anti-E. coli LPS antibody [2D7/1]

     Excellent
    Abreviews
    abreview image
    Application
    ELISA
    Sample
    Mouse Serum (Intestinal fluid)
    Specification
    Intestinal fluid
    Type
    Sandwich (Capture)
    Blocking step
    Pierce No Protein Blocking Buffer as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 100% · 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.

    Sarah Hoyle

    Verified customer

    Submitted Nov 12 2013

    Question

    I have a few questions relating to the abovementioned product number.

    1) Was the animal immunised with whole cell J5 or LPS from J5 cells ? If whole cells were injected then where does the LPS specificity come from. Did the manufacturers do epitope mapping?

    2) Is the antibody E coli J5 specific or does it detect across a range of E coli serotypes. Was cross reactivity studies conducted by the manufacturer and if so, are these results available ?

    We are looking at purchasing a broad spectrum Anti E coli antibody and therefore clarity on these questions are important before a decision is made to purchase.

    Read More
    Answer

    Thank you for contacting us and your interest in our products.

    The Anti-E. coli LPS antibody [2D7/1], ab35654, has been raised against whole E. coli cells (rather than disrupted or sonicated cells). Whilst specific epitope mapping has not been performed, the outer cell wall of E. coli is predominantly LPS and one would need to remove this before the other cell wall proteins would be exposed. Additionally, information from the developers of this clone have indicated that the antibody does indeed detect J5 LPS by ELISA. Unfortunately, cross reactivity with other E. coli proteins has not been performed so this does remain a possibility (albeit a remote one).

    Aside from E. coli J5, the clone was found to be reactive to K. pneumoniae, S. sonnei and S. typhimurium. Unfortunately, it has not been tested against other E. coli serotypes and we currently do not have any empirical data to indicate which other serotypes, if any, are recognised by this clone.

    I hope this has been helpful. If you have any further questions, please do not hesitate to let me know.

    Read More

    Question

    Phone call requesting information on if the antibody is likely to corss react with E. coli J 53 stain

    Read More
    Answer

    Thank you for contacting us and your interest in the Anti-E. coli LPS antibody [2D7/1] (ab35654).

    I now have further information to share in regards to how this antibody has been investigated for cross reactivity with LPS from other strains than the E. coli J5 strain. Unfortunately, we have not specifically tested this antibody on the J53 strain. However, the clone is known to also react to the following species:

    K. pneumoniae, S. sonnei and S. typhimurium

    Given this, I believe it is likely to react to other E.coli strains as well.

    I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

    Read More

    Question

    Hi,

    What is the cat# of your LPS (or is it the strain that is available in your catalugue?). I need a LPS and an ab against LPS that I know would work so if you sell LPS that you know works I would love to buy some. I guess that I could also try the Abreview aswell sice I have the other LPS here.

    I will order it as soon as I get the cat#, I could not find it in your catalogue.

    All the best

    Read More
    Answer

    Thank you for your reply.

    Unfortunately, we currently only have Pichia pastoris LPS protein (ab73792) available from our catalogue. However, we do not have an anti-Pichia pastoris LPS antibody available, I cannot therefore suggest a suitable pair for you.

    If you would be interested in the Abtrial testing discount scheme discussed in my previous email please do let me know and I can raise a discount code for you. More information on this offer can be found from the following link:

    www.abcam.com/abtrial

    I am sorry that I could not be of more help in this instance. If you have any further questions, please do not hesitate to contact us again.

    Read More

    Question

    Product code: 35654

    Inquiry: Hi, I am looking for an anti LPS ab. I have LPS from xxxx) made from e.coli strain 026:b6. My question which of your antibody would you recommend me to buy from you? If no suggestion could be made, would it then be possible to get a smal sample of your ab35654 to try it out. Thanks for all the help

    Read More
    Answer

    Thank you for contacting us and your interest in our products.

    The anti-E. coli LPS antibody [2D7/1], product code (ab35654), was raised against the J5 strain of E.coli, which produces incomplete LPS. The product you have purchased from xxxx is from a different strain of E.coli with short chain length LPS. The ab35654 has not been epitope mapped or tested for reactivity against LPS from strain O26:B6. So unfortunately we could not guarantee it’s reactivity for use with the your LPS, although it may well bind. The O26:B6 strain produces similar LPS to that of the mutant rough chain LPS from the J5 strain.

    Unfortunately, we currently only have antibodies raised against the J5 strain LPS currently available from the catalogue. Due to the size of our catalgoue we are not able to provide sample sizes of any of our products. It simply is not something that we can logistically perform. I would however you would be able to participate in the Abtrial.

    This offer means that if you purchase ab35654 as normal, test the antibody with your samples and let us know of the results through an Abreview (no matter whether positive or negative) you would be eligible for the equivalent price of ab35654 off any product in our catalogue. More information on this offer can be found here:

    https://www.abcam.com/abtrial

    Please note that the antibody to be tested must be purchased, tested, the Abreview submitted and the free product claimed within a 4 month period.

    If you would be interested in participating in this scheme please do let me know as a discount code needs to be issued prior to the purchase of ab35654.

    I hope this information has been of some help. If I can be fo any further assistance, please do not hesitate to contact us again.

    Read More

    Question



    Inquiry: Hello, We used your Anti-E. coli LPS antibody [2D7/1] to detect LPS on E. Coli however using epifluorescence microscopy, we were not able to detect any signal. We first fixed two different strains of E. coli (J5 and o111, a pellet from 50ul of overnight growth ) with 4% paraformaldehyde (in 1X PBS). The fixed pellets were then each resuspended in 100ul 1X PBS. We then added a concentration of 1:100 of the anti-LPS to each strain, and incubated the samples for 30 min at room temperature. Each strain was then washed 3 times with 500ul of 1X PBS and then a 1:100 concentration of the secondary antibody AF 488 rabbit, anti-mouse was added to each sample and incubated for 30 minutes. Each sample then was washed 3 times with 1X PBS and then placed on a slide and viewed under the microscope. We also made controls of secondary AF 488 only, a isotype IgG control, and an unstained sample, and were not able to detect any signal for any of the samples. We were wondering if you could tell us why we were not able to detect LPS signal, suggest any alternate way we could use to verify that the anitbody works, as well as if fixing the E. coli strains with paraformaldehyde affected the LPS-antibody interaction? Thank you

    Read More
    Answer

    Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

    The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

    Having reviewed this case, I am curious about the possibility that the secondary antibody is causing the problem. You state you used the IgG control and yet you did not any staining, which indicates it is not the primary antibody. Do you have another secondary antibody which you could try?
    Should the suggestions not improve the results, please do let me know.

    In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

    I hope this information is helpful, and I thank you for your cooperation.

    Read More

    Question

    Hello,
    Thank you for your reply. Would it be possible to have a few microliters of the antiserum to test it in our tissue please?
    Regards,

    Read More
    Answer

    Thank you for contacting us. Because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products.

    We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

    I am happy to apply the Abpromise guarantee on this product in the case it is not working on the strain you are studying.

    I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

    Read More

    1-10 of 32 Abreviews or Q&A

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