Overview

  • Product name
    Goat Anti-Chicken IgY H&L (HRP) preadsorbed
    See all IgY secondary antibodies
  • Host species
    Goat
  • Target species
    Chicken
  • Tested applications
    Suitable for: WB, Immunomicroscopy, ICC/IF, Dot blot, ELISA, IHC-P, IHC-Frmore details
  • Minimal
    cross-reactivity

    Cow, Goat, Guinea pig, Hamster, Horse, Human, Mouse, Rabbit, Rat, Sheep more details
  • Immunogen

    Full length native chicken IgY (purified).

  • Conjugation
    HRP

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Gentamicin sulphate
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride, 1% BSA
  • Concentration information loading...
  • Purity
    Affinity purified
  • Purification notes
    This product was prepared from monospecific antiserum by immunoaffinity chromatography using chicken IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities.
  • Conjugation notes
    Horseradish Peroxidase (HRP)
  • Clonality
    Polyclonal
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab7118 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
WB Use at an assay dependent dilution.
Immunomicroscopy Use at an assay dependent dilution.
ICC/IF 1/1000 - 1/2500.
Dot blot Use at an assay dependent dilution.
ELISA 1/20000 - 1/100000.
IHC-P Use at an assay dependent dilution.
IHC-Fr Use at an assay dependent dilution.

References

ab7118 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Thank you for your quick response.

The Questionnaire has been attached to my first response as a word document but I have reattached it to this e-mail as well.

Please could you add further information to the Questionnaire? I am particularly interested in sample preparation, lysis buffer. Have you run a secondary antibody ONLY control (without the sample and primary antibody)? Liver cells/tissues have very high concentration of intracellular proteins. It may be worth increasing the concentration of the blocking agent from 5-10%. Have you had any chance to use BSA rather than milk for blocking?

Could you please confirm the Abcam Order number and the arrival date as well?

I look forward to hearing from you soon.

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Question

1) Abcam product code ab7118



2) Abcam order reference number or product batch number lot nr GR27997-6



3) Description of the problem: unspecific bands on western, with using only the secondary antibody (no primary)



4) Sample preparation:

Type of sample (whole cell lysates, fraction, recombinant protein…): tissue homogenates, centrifuged 4000xg, supernatants dissolved in Laemli

Lysis buffer: Laemli (SDS, DTT)

Protease inhibitors: Pefabloc (SDigma-Aldrich) + Leupeptin

Phosphatase inhibitors: no

Reducing agent: DTT

Boiling for ≥5 min? yes/no: 65C 15 min

Protein loaded ug/lane or cells/lane: approximately 10-30 ug protein per lane

Positive control

Negative control



5) Percentage of gel: 4-15% polyacrylamide

Type of membrane: PVDF

Protein transfer verified: the marker, which is blue is gone from the gel, and can be seen on the membrane

Blocking agent and concentration: milk 5%

Blocking time: 1 h

Blocking temperature: RT



6) Primary antibody (If more than one was used, describe in “additional notes”) :

NO PRIMARY ANTIBODY!



7) Secondary antibody:

Species: goat

Reacts against: chicken

Concentration or dilution: 1:5000

Diluent buffer: PBS-T

Incubation time: 1 h

Incubation temperature: RT

Fluorochrome or enzyme conjugate: HRP



8) Washing after primary and secondary antibodies:

Buffer PBS-T

Number of washes: 4-5



9) Detection method: ECL



10) How many times have you run this staining? Two times, with different samples, because I thought first that something was wrong with my samples.

Do you obtain the same results every time? yes

What steps have you altered to try and optimize the use of this antibody? Not optimized, I would like hear first what your suggestions are.



Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.







From: technical@abcam.com [mailto:technical@abcam.com]
Sent: 12. oktober 2012 09:32
To: Aleksandra Rojek
Subject: Reply from Abcam to your enquiry regarding ab7118 [CCE4241379]



















https://www.abcam.com/index.html?utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Header









Dear Dr Rojek

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).

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.




Best regards,
Tanya

Tanya Bagrij, PhD
Scientific Support Supervisor
Abcam plc
www.abcam.com


Your original inquiry to Abcam:


Lot number: GR27997-6

Enquiry: Hi, I experience problems with the secondary antibody: I see bands on the blot, without adding primary antibody. This would indicate unspecific binding of secondary antibody to the proteins on the membrane. How do I avoid unspecific binding of secondary ab? Is there any way of more stringent washing (protocol)? WB conditions: 20 ul mouse liver homogenate per lane, blocking 5% milk, detection ECL, 4-15% polyacrylamide gel, dry transfer, secondary 1:5000 diluted in PBS-T 1hr RT, washes 4-5 times 5-15 min. (This protocol worked fine previously with different primary ab made in rabbit, and goat anti rabbit secondaries, and we have not previously experienced unspecific signals from secondary alone) Best regards,




Abcam Customer Services and Scientific Support Team
https://www.abcam.com/index.html?pageconfig=technical&utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Body

[CCE4241379]













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Answer

Thank you for your enquiry regardingab7118 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

Though you have kindly provided some details, it would be much appreciated if I could get some more information which would help me identify the source of the problem.

- Have you run a secondary antibody ONLY control (without the sample and primary antibody)?

- Liver cells/tissues have very high concentration of intracellular proteins. Have you tried loading 5 or ug per lane?

- It may be worth increasing the concentration of the blocking agent from 5-10%. Have you had any chance to use BSA rather than milk for blocking?

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

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).

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

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

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