Overview

  • Product name
    Anti-E. coli LPS antibody [2D7/1]
  • Description
    Mouse monoclonal [2D7/1] to E. coli LPS
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-Fr, ICC/IF, IHC-P, IP, ELISA, WBmore details
  • Species reactivity
    Reacts with: Escherichia coli
  • Immunogen

    Tissue/ cell preparation: Escherichia coli J5 cells.

Properties

Applications

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
IHC-Fr Use at an assay dependent concentration.
ICC/IF 1/200.
IHC-P Use at an assay dependent concentration.
IP Use at an assay dependent concentration. PubMed: 18604629
ELISA 1/1500.
WB Use at an assay dependent concentration. PubMed: 22492977

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

  • 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

  • ab35654 staining E. coli LPS in rat intestinal tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    Tissue was fixed in formaldehyde and a heat mediated antigen retrieval step was performed using citrate EDTA buffer pH 6.2. Samples were then blocked and incubated with ab35654 at a 1/100 dilution for 1 hour. The secondary used was a polyclonal goat anti-mouse IgG conjugated to alkaline phosphatase, used at a 1/100 dilution.

    See Abreview

References

This product has been referenced in:
  • Carnevale R  et al. Localization of lipopolysaccharide from Escherichia Coli into human atherosclerotic plaque. Sci Rep 8:3598 (2018). Read more (PubMed: 29483584) »
  • Cañas MA  et al. Outer Membrane Vesicles From Probiotic and Commensal Escherichia coli Activate NOD1-Mediated Immune Responses in Intestinal Epithelial Cells. Front Microbiol 9:498 (2018). ICC/IF ; Human . Read more (PubMed: 29616010) »
See all 23 Publications for this product

Customer reviews and Q&As

1-10 of 32 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Dec 11 2018

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

Catherine Brennan

Verified customer

Submitted Jan 10 2018

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

Abcam user community

Verified customer

Submitted May 19 2015

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

Sarah Hoyle

Verified customer

Submitted Nov 12 2013

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

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.

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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

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

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

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.

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1-10 of 32 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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