山羊抗鸡IgY H&L (HRP)预吸附二抗(ab7118)

Submit a review Q&A (3)

Key features and details

  • Goat Anti-Chicken IgY H&L (HRP) preadsorbed
  • Conjugation: HRP
  • Host species: Goat
  • Suitable for: WB, Immunomicroscopy, ICC/IF, Dot blot, ELISA, IHC-P, IHC-Fr

Conjugates logo Related conjugates and formulations

Agarose Alexa Fluor® 405 Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 680 Alexa Fluor® 750 Alexa Fluor® 790 Alkaline Phosphatase Biotin Cy2 ® Cy3 ® Cy5 ® Cy5.5 ® DyLight® 488 DyLight® 550 DyLight® 594 FITC Gold 10nm Gold 25nm HRP PE Texas Red ® Unconjugated Unconjugated

概述

  • 产品名称

    山羊抗鸡IgY H&L (HRP)预吸附二抗
    参阅全部 IgY 二抗

  • 宿主

    Goat

  • 靶标种属

    Chicken

  • 经测试应用

    适用于: WB, Immunomicroscopy, ICC/IF, Dot blot, ELISA, IHC-P, IHC-Frmore details

  • 最小交叉反应


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

  • 免疫原

    Full length native chicken IgY (purified).

  • 偶联物

    HRP

性能

  • 形式

    Liquid

  • 存放说明

    Shipped at 4°C. Store at +4°C.

  • 存储溶液

    Preservative: 0.01% Gentamicin sulphate
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride, 1% BSA
  • Concentration information loading...

  • 纯度

    Affinity purified

  • 纯化说明

    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.

  • 共轭说明

    Horseradish Peroxidase (HRP)

  • 克隆

    多克隆

  • 研究领域

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab7118于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
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.
说明
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.

实验方案

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

文献 (0)

发表研究结果有使用 ab7118？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab7118 尚未被引用在任何文献中。

客户评价及客户问答

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1-3 of 3 Abreviews or Q&A

Question

Where is the questionnaire that I should fill out? If it is the note with product number and lot number, I already filled that out in my previous inquiry.

Best regards,

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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.

Read More

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

Question

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,

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

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