Overview

  • Product name
    Anti-E. coli antibody (HRP)
    See all E. coli primary antibodies
  • Description
    Rabbit polyclonal to E. coli (HRP)
  • 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.

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

This product has been referenced in:
  • Sanvicens N  et al. Quantum dot-based array for sensitive detection of Escherichia coli. Anal Bioanal Chem 399:2755-62 (2011). Read more (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). Read more (PubMed: 18256810) »
See all 6 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Question
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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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

Read More

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

Read More
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.

Read More

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.

Read More

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

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