Overview

  • Product name
  • Description
    Goat polyclonal to E. coli
  • Host species
    Goat
  • Specificity
    ab25823 recognises all E. coli species tested. The following species of E. coli were used in the antigen prep: Detergent lysed E. coli from K12, TOP 10F, XS127/P3, JM109,HB101, and DH5a and BL21. We have also extensively tested K-12 Since there is 95% homology between the strains, all species of E.coli should react with this antibody.
  • Tested applications
    Suitable for: ICC/IF, Electron Microscopy, WB, ELISA, Dot blotmore details
  • Species reactivity
    Reacts with: Escherichia coli
  • Immunogen

    Tissue, cells or virus corresponding to E. coli. Tissue/ cell preparation (Escherichia coli ): Detergent lysed E. coli (cocktail blend of strains of E. coli including the most commonly used DH5a and BL21).

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Constituents: 0.595% HEPES, 0.87% Sodium chloride

    The buffer contains trace amounts of citric acid and sodium phosphate. There are no preservatives or any other amine containing compounds and as such this material can be directly conjugated to enzymes, biotin etc. by most chemistries without the need to diafilter.
  • Concentration information loading...
  • Purity
    Protein G purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab25823 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 Use at an assay dependent concentration. PubMed: 22427638
Electron Microscopy Use at an assay dependent concentration. PubMed: 24253282
WB Use a concentration of 0.5 - 2 µg/ml. Note: Reacts with more than 50 proteins by Western Blot.
ELISA Use at an assay dependent concentration.
Dot blot 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:
  • Farka Z  et al. Rapid detection of microorganisms based on active and passive modes of QCM. Sensors (Basel) 15:79-92 (2014). Read more (PubMed: 25545267) »
  • Nayak M  et al. Integrated sorting, concentration and real time PCR based detection system for sensitive detection of microorganisms. Sci Rep 3:3266 (2013). Electron Microscopy ; Escherichia coli . Read more (PubMed: 24253282) »
See all 6 Publications for this product

Customer reviews and Q&As

1-10 of 21 Abreviews or Q&A

Abreviews
Application
ELISA
Sample
Escherichia coli Cell (K12 bacterial solution)
Specification
K12 bacterial solution
Type
Other
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1%

Abcam user community

Verified customer

Submitted May 13 2014

Answer


The Goat polyclonal anti-E. coli conjugated to HRP is ab69185. The link to the datasheet is https://www.abcam.com/ab69185.

Read More

Answer

Thank you for contacting us.

We only supply this antibody in 100 ug sizes however, if you were to purchase 5 or more vials of this product, it would be possible for me to give you a discount.

If you could let me know how many vials you would like to purchase from us, I will then enquire about the discount that we can offer you.

I look forward to your reply.

Read More

Answer

Thank you for contacting us.


The K12 strain was one of the strains used to generate the antibody.

This antibody was actually raised against a blend of lysates from both BL21 and DH5a and as a result will detect the majority of ECPs as evidenced by 1 dimensional western blotting. Because most ECP antigens are conserved between various strains (>90% homology, most likely determined by sequence alignment), this antibody has proven useful in detection ECPs from a great variety of E.coli strains including but not limited to the following; K12, JM109, TOP 10F HB101, and ATCC 10798. We also know that this antibody will recognize the strains O18, O20, O44, O55, O111, O112, O125, O157.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Question
Answer

Thank you for contacting Abcam.

The SDS for ab25823 has been created and will be posted on the website by the "Datasheet PDF" tab within the next 48 hours (https://www.abcam.com/e-coli-antibody-ab25823.html).

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Answer

Thank you for your email.

I understand your concerns that antibodies should form immune complex with compatible antigens irrespective of their location however unfortunately this is not the case. Antibodies behaves differently in each application and with experimental conditions. It may well be that the problem of enrichment you are experiencing is because of antibody being not binding to magnetic beads. I am very sorry we do not guarantee our antibodies for this.

Secondly, I agree that we will be happy in replacing the antibody if it fails in ELISA. Please do contact me with results once your finish your experiment.

I hope this information will be helpful. Should you have any other question please do not hesitate to contact me.

Read More

Question

Below you may find the abcam questionaire. Since that I used this antibody to immobilize on magnetic particles for immunomagnetic separation of E.coli, I adapt this questionaire for an immunomagnetic separation application.
Best regards,
Delfina
1) Abcam product code ab ab 25823

2) Abcam order reference number or product batch number: 1134800

3) Description of the problem: No E.coli captured; The antibody does not recognize E.coli in medium

4) Type of application: Immunomagnetic separation

5) Sample details: E.coli (DH5a, MC 1061 and CGSG 5073) in BHI medium

6) Target antigen (if found in heterogeneous sample)

7) Positive control: N/A
8) Negative control: BHI medium

9) Blocking agent (eg BSA…): BSA to block the magnetic particles with immobilized ab 25823
Concentration: 0.5 %
Blocking time: 2h
Blocking temperature: 37 ºC

10) Capture or Primary antibody information: ab 25823
Used to coat wells? no
Species: goat
Reacts against ab25823 recognises all E. coli species tested. The following species of E. coli were used in the antigen prep: Heat killed sonicate of E. coli from K12, TOP 10F, XS127/P3, JM109,HB101, and DH5a and BL21. We have also extensively tested K-12 Since there is ]95% homology between the strains, all species of E.coli should react with this antibody.
Concentration or dilution: 1 mg/ mL
Diluent buffer : PBST
Incubation time: 30 and 1h
Incubation temperature: RT

11)Detection antibody: N/A
Species:
Reacts against
Concentration or dilution
Diluent buffer
Incubation time
Incubation temperature
Conjugation
12) Enzyme labelled secondary antibody: N/A
Species
Reacts against
Concentration or dilution
Diluent buffer
Incubation time
Incubation temperature:
Conjugation
13) Washing after primary and secondary antibodies:
Buffer: PBST
Number of washes: 3
Length of washes: 1 min
14) Substrate:N/A
Incubation time:
15) How many times have you run this application? 3
Do you obtain the same results every time? yes
What steps have you altered to try and optimize the use of this antibody? Increase the antibody concentration and compared with different concentrations and strains of E.coli.

Read More
Answer

Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

I have read the details you have kindly provided and have following further questions for better understanding of the problem;

- What type of Dynabeads you have used e.g. Protein A or Protein G coupled or both?
- How long you have incubated the antibody with Dynabeads?
- Have you done any troubleshooting in incubation timing of Dynabeads with antibody and sample? Please provide details.
- Is the conjugation with direct or indirect method?
- Is the blocking step with BSA recommended by the manufacturer of Dynabeads?
- You have mentioned observing 20% capture efficiency with antibody. How the efficiency was tested? Was it with IPed samples?
- Is the antibody used in any other application?

Please note, ab25823 has not been tested in IP, which means the use of product in IP is out of Abpromise guarantee so we will not be able to replace the product. We however will help you in troubleshooting for achieving best results.

Thank you very much for your cooperation. I will 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.

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 contacting us.

I have heard back from the lab with the following answers:

1. ab25823 can be used for Western blot in the concentration range of 0.5 to 2ug/mL.

2. We have not tested the O157 strain for cross reactivity but a previous customer has claimed that it does. We would expect there are many proteins that are conserved among various E.coli strains but we cannot put a number on how many O157 proteins react with ab25823.



But if you like to try the antibody in the O157 strain, you can participate in our testing discount program.

For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:

The testing discount program is for customers who like to use an antibody/protein/kit on an untested species/application. You would purchase the antibody/protein/kit at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody/protein at the full price (100%) of the antibody/protein you have tested, or a full price discount for the amount paid for the kit. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.
This programapplies to ELISA kits and other kits we offer.

Please let me know if you like to receive a testing discount code.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

Merci de nous avoir contactés.
Comme expliqué par téléphone j'ai mis en pièce jointe un questionnaire qui nous permettra de regarder plus en détails votre protocole.
Les liens vers les protocoles Abcam pour la cytométrie en flux sont:
> https://www.abcam.com/index.html?pageconfig=resource&rid=11381
> https://www.abcam.com/index.html?pageconfig=resource&rid=11443
N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

Read More

1-10 of 21 Abreviews or Q&A

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

Sign up