HRP Anti-E. coli antibody (ab20425)

Submit a review Q&A (5)References (6)

Key features and details

  • HRP Rabbit polyclonal to E. coli
  • Suitable for: ELISA, ICC/IF
  • Reacts with: Escherichia coli
  • Conjugation: HRP
  • Isotype: IgG

Overview

  • Product name

    HRP Anti-E. coli antibody
    See all E. coli primary antibodies

  • Description

    HRP Rabbit polyclonal to E. coli

  • Host species

    Rabbit

  • Conjugation

    HRP

  • Specificity

    All O and K antigenic serotypes of Escherichia coli. Antiserum is not absorbed and does react with related Enterobacteriaceae.

  • Tested applications

    Suitable for: ELISA, ICC/IFmore details

  • Species reactivity

    Reacts with: Escherichia coli

  • Immunogen

    Mixture of E.coli serotypes

  • General notes


    Will remove E. coli proteins from recombinant preparations.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C.

  • Storage buffer

    Preservative: 0.002% Thimerosal (merthiolate)
    Constituents: PBS, 1% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Primary antibody notes

    Will remove E. coli proteins from recombinant preparations.

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab20425 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
ELISA 1/2000.

Against serotypes O157:H7, O20, O125, O55, O111 and K12.
ICC/IF Use at an assay dependent concentration.

Target

  • Relevance

    Escherichia coli is a gram negative bacillus that belongs to a larger group of Enterobacteriae - bacteria that inhabit the gastrointestinal tract. Although usually a harmless resident of the gut, some strains have the potential to cause serious problems, especially where there is an immature immune system or immunosuppression, or where the subtype of organism has acquired the ability to produce pathogenic toxins.

  • Alternative names

    • Escherichia coli antibody

References (6)

Publishing research using ab20425? Please let us know so that we can cite the reference in this datasheet.

ab20425 has been referenced in 6 publications.

  • Sanvicens N  et al. Quantum dot-based array for sensitive detection of Escherichia coli. Anal Bioanal Chem 399:2755-62 (2011). PubMed: 21249344
  • Baldrich E  et al. Self-assembled monolayers as a base for immunofunctionalisation: unequal performance for protein and bacteria detection. Anal Bioanal Chem 390:1557-62 (2008). PubMed: 18256810
  • Baldrich E  et al. Gold immuno-functionalisation via self-assembled monolayers: study of critical parameters and comparative performance for protein and bacteria detection. J Immunol Methods 336:203-12 (2008). ELISA ; Escherichia coli . PubMed: 18534611
  • Laczka O  et al. Immunofunctionalisation of gold transducers for bacterial detection by physisorption. Anal Bioanal Chem 391:2825-35 (2008). ELISA ; Escherichia coli . PubMed: 18553187
  • Lin YH  et al. Disposable amperometric immunosensing strips fabricated by Au nanoparticles-modified screen-printed carbon electrodes for the detection of foodborne pathogen Escherichia coli O157:H7. Biosens Bioelectron 23:1832-7 (2008). PubMed: 18424027
  • Laczka O  et al. Detection of Escherichia coli and Salmonella typhimurium using interdigitated microelectrode capacitive immunosensors: the importance of transducer geometry. Anal Chem 80:7239-47 (2008). PubMed: 18771278

Customer reviews and Q&As

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1-5 of 5 Abreviews or Q&A

Question

please send credit note.

thank you

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Answer

This e-mail was sent to you on the 20th September.

CCEI: http://ops.adminsite.com/admin/crm2/crm.cfm?mid=4167113

Thank you for contacting us.

Your credit note ID is 22726.

I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at mailto:creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

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Question

From what I can see on the "quittung" I have, I would say we got it from LUCERNA CHEM AG.
Could it be helpful?

Thank you very much and sorry for the complication!

Best Regards

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Answer

Thank you for contacting us.

Your credit note ID is 22726.

I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

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Question

Thank you very much for asking, I was under a mountain of things to do and forgot to send the reply.

I don't find the form you sent me, so I quickly write all the parameters her, together with graphs:

Briefly, left graph shows the signal obtained from a routine test we developed with a custom primary Ab, and a secondary Ab-HRP coupled. The substrate was ABTS ready solution from thermo scientific. Blue line is positive control (bacteria with Type 1 pili and expressing a protein we use to trap them on the wells), red negative control (same bacteria, with same pilus, but no adhesion protein). Primary Ab is polyclonal against the pilus. They are captured through the adhesion protein.

Central graph is the result of a test run same way. Instead of primary and secondary Ab, I wanted to use a general E.coli Ab, so that in one single step I would have been able a similar result. Again, blu is positive, red is negative.

Third graph is a sensibility comparison, so both lines are for the positive, and show that our in-house system is 4 time more sensible, under same conditions.

As you can see, I tried dilution 1:500 - 1:4000 of the Ab, so a quite broad range.

The protocol:

1.coating overnight at 4° our trap element (can't say more)

2.wash twice with PBS, then blocking 2h with 1% BSA in PBS (commercial, from Nalgene). strategy tested also for our system, it works very well.

3.wash 2x with PBS.

4.incubation with E.coli 2h

6.3x wash withphosphate buffer saline 100mM K2HPO4, 150mM NaCl, containing tween 20 0.05%. 5 minutes each wash with shaking

5.incubation with primary Ab 2h

6.wash 3x with phosphate buffer saline 100mM K2HPO4, 150mM NaCl, containing tween 20 0.05%. 5 minutes each wash with shaking

7.incubation with secondary Ab 2h

8. 3x wash as in 6

9.development ABTS (in the dark, covered with foil)

The protocol in case of your Ab a-E.coli-HRP:

1.coating overnight at 4° our trap element (can't say more)

2.wash twice with PBS, then blocking 2h with 1% BSA in PBS (commercial, from Nalgene). strategy tested also for our system, it works very well.

3.wash 2x with PBS.

4.incubation with E.coli 2h

5.3x wash withphosphate buffer saline 100mM K2HPO4, 150mM NaCl, containing tween 20 0.05%. 5 minutes each wash with shaking

6.incubation with Ab-HRP 2h

7.wash 3x with phosphate buffer saline


8.development ABTS(in the dark, covered with foil)

As you can see from the graph, the Ab is not sensible, also at high concentration. It recognizes the wells in which there is E.coli, but a signal is visible only for high OD, and the signal to background ratio is very bad.

As you can see from the absorbance value, this is not a problem of high background. It is in fact very low, and in the same range of our in-house test.

The impression I had, is that this Ab, although should be general, is not really efficient in recognizing our strain.

Thank you very much for the assistance,

Best Regards

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Answer

Thank you for getting back tome promptly and for providing some useful details.

As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

Would you be so kind to confirm if these experiments have been carried out head-to-head or separately. It may well be that either ab20425 has been accidentally damaged during shipment/storage the affinity is slightly different from the other product.

Since you have an alternative product which provided the expected results, I could offer you a credit note which you can use in future purchase.

Coudl you please confirm your Abcam Order Number (or PON) and the date of purchase?

Let me know how you wish to proceed with your enquiry.

I look forward to hearing from you soon.

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Question

Product code: 20425
Lot number: gr74821-1
Inquiry: Dear Sirs, I purchased an antibody from you and I am having some troubles. The code is ab20425. It should work against all O and K antigenic serotypes of E.Coli. I used it against UTI89 E.Coli, but the Ab seems not to be specific for them. Is it possible that I made some mistakes, or that this serotype is not recognized? Thank you for your help, Best Regards 96-well plate coated with a standard methodology a mannosylated protein Incubation with different bacteria dilutions (from1billion to 10000cfu/ml) washing steps with PBS incubation with Ab 20min 1:2000 and 1:200 washing steps with PBS ABTS substrate color development: no difference among the different dilutions. Negative control (same procedure, no bacteria) showed comparable color.

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Answer

I am sorry to hear that you have been experiencing problems using this product in the application that you wish.

In order to assess the quality of our products I would ask that you complete a brief questionnaire relating to the application used. Often it is possible to make suggestions that may help resolve problems experienced using a particular product.

As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

All our customer feedback, including complaints are monitored weekly by our in house technical support team. If a product is at fault the technical support team will consider removing the product from our catalogue in order to avoid future customer inconvenience.

Please could you provide some further details of the protocol used and complete the following form (attached as a word document). It would be much appreciated if you could attach an image to the response.

Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

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Question

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Answer

Thank you for your enquiry. Further to our telephone conversation this afternoon I have performed a detailed search of our catalogue for antisera that recognizes E.coli subclass O127. We do not have an antibody that uniquely recognizes this serotype. However, antibodies ab13626, ab20425 and ab20640 all recognise all O antigenic serotypes of Escherichia coli. ab20640 is conjugated to Biotin. I hope this information helps, please do not hesitate to contact me should you require further assistance.

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