山羊F(ab')2抗小鼠IgG - H&L (HRP),预吸附二抗(ab5870)

Submit a review Q&A (6)References (3)

Key features and details

  • Goat F(ab')2 Anti-Mouse IgG - H&L (HRP), pre-adsorbed
  • Conjugation: HRP
  • Host species: Goat
  • Isotype: IgG
  • Suitable for: ICC/IF, Dot blot, WB, ELISA, Electron Microscopy, IHC

概述

  • 产品名称

    山羊F(ab')2抗小鼠IgG - H&L (HRP),预吸附二抗
    参阅全部 IgG 二抗

  • 宿主

    Goat

  • 靶标种属

    Mouse

  • 经测试应用

    适用于: ICC/IF, Dot blot, WB, ELISA, Electron Microscopy, IHCmore details

  • 最小交叉反应


    Cow, Horse, Human, Rabbit, Rat, Sheep more details

  • 免疫原

    Mouse IgG whole molecule.

  • 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

    pH: 7.20
    Preservative: 0.01% Gentamicin sulphate
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride, 1% BSA

    BSA is IgG and protease free
  • Concentration information loading...

  • 纯化说明

    This product was prepared from monospecific antiserum by immunoaffinity chromatography using Mouse IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities, pepsin digestion and chromatographic separation.

  • 共轭说明

    Horseradish Peroxidase (HRP) (Molecular Weight 40,000 daltons)

  • 克隆

    多克隆

  • 同种型

    IgG

  • 研究领域

应用

Our Abpromise guarantee covers the use of ab5870 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF 1/500 - 1/2500.
Dot blot Use at an assay dependent dilution.
WB 1/1000 - 1/10000.
ELISA 1/60000.
Electron Microscopy Use at an assay dependent dilution.
IHC 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 (3)

发表研究结果有使用 ab5870？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab5870 被引用在 3 文献中.

  • Morash AJ  et al. Pass the salt: physiological consequences of ecologically relevant hyposmotic exposure in juvenile gummy sharks (Mustelus antarcticus) and school sharks (Galeorhinus galeus). Conserv Physiol 4:cow036 (2016). PubMed: 27757235
  • Tunnah L  et al. Physiological responses to hypersalinity correspond to nursery ground usage in two inshore shark species (Mustelus antarcticus and Galeorhinus galeus). J Exp Biol 219:2028-38 (2016). WB ; Shark . PubMed: 27207636
  • Ulasov IV  et al. Survivin-driven and fiber-modified oncolytic adenovirus exhibits potent antitumor activity in established intracranial glioma. Hum Gene Ther 18:589-602 (2007). WB . PubMed: 17630837

客户评价及客户问答

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Question

I am looking for a HRP conjugated anti-mouse antibody to be used in ELISA. The most important thing is that it will have the least cross reactivity to rat immunoglobulins. I see that you have many such antibodies. Which one would you think is the most suitable?


Thanks,

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Answer

Thank you for contacting us.

The following products would be suitable as per your requirements. The catalogue numbers are

ab5870, ab5887, ab7061, ab7068, ab98717, ab98790 etc

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

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Question

Thank you for your information in the previous email. I have an enquiry about the secondary antibody-HRP. What's the difference between ab5887 and ab5870?

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Answer

Thank you for your question. Both antibodies are only a F(ab)2 Fragment of antibody itself (therefore these antibody will not give F(c) mediated binding), but they different in the immunogens used and therefore epitopes recognized: Ab5887 was raised against Mouse IgG F(ab)2 fragment. Ab5870 was raised against Mouse IgG whole molecule. I hope this information will help. Please do not hesitate to contact us if you need further assistance.

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Question

May I ask once more? My samples are sheep proteins. I am going to use two antibodies (anti GAPDH and anti GDF8 antibodies). The first antibodies are raised in mouse and rat (mouse IgG and Rat IgG), respectively. There are many secondary antibodies (HRP conjugated) which recognize mouse IgG or rat IgG (ex. ab5870, ab5879, ab5887, ab6728.....). As some of them are raised in goat, I am afraid that I will have a high background. I would like to know which secondary antibodies against mouse and rat IgG is good for western blotting to detect sheep proteins. Thank you for your helps.

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Answer

Thank you for your enquiry. Given that you are using mouse and rat primary antisera and would like to use a secondary antiserum that will not detect any endogenous IgG in your sheep samples, bearing in mind a potential cross reactivity with goat, I would like to recommend the following: ab6728 Mouse IgG antibody (ab6728) HRP conjugated Rabbit ab6820 Mouse IgG antibody (ab6820) HRP conjugated Donkey Both antisera have been raised in rabbit or donkey and are highly unlikely to cross react against sheep. We have also received favourable reviews for them both.

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Question

Thank you for your info. As I mentioned in my email, I used the same kind secondary antibody from Pierce to compare with your product. It showed a nice signal at dilution 1:20000 whereas your product showed a similar signal at dilution 1:100 under the same experiment condition! I bought your product for a direct ELISA, but it didn't work. That's the reason why I did this control test to coat primary antibody directly to the plate. If you believe it just a bad vial, send me a replacement and I will try the test again.

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Answer

Thank you for your clarification and for sharing more details with us. Certainly, we can offer you a replacement vial - free of charge. Would you be so kind to confirm your Purchase Order Number, or Abcam Order Number. We look forward to hearing from you soon.

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Question

1. Please describe the problem (high background, no signal, non-specific colour development, poor standard curve etc). Signal is weaker than normal antibody. 2. What type of ELISA are you performing? (Direct ELISA, indirect ELISA, sandwich ELISA etc.) Direct ELISA. 3. On what material are you testing the antibody in ELISA? Species? Cell type? etc? Coat the plate with primary Antibodies. 3. How did you coat the plates? Coating buffer: 20mM Tris-HCL pH8.5 100ul/well, primary Ab 1:1000. 4. How did you block the unspecific binding sites? For how long? How did you wash the wells? 3% BSA in TBS, 4 hours, wash by TBST buffer 5X between each step. 5. Primary antibody Specification (in which species was it raised against)? rat anti-HA, mouse anti-BDG At what dilution(s) have you tested this antibody? 1:1000, 1:500 Incubation time, wash steps (multiple short washes are more effective than fewer longer wash steps)? 2 hours, 5X TBST 6. Secondary antibody What secondary antibody are you using? Your ab6120-250, ab5870-250. Specification (in which species was it raised against)?goat At what dilution(s) have you tested this antibody? Incubation, wash steps? 1 hour, 5XTBST Do you know whether the problems you are experiencing come from the secondary? I also used second antibody from Pierce as a control, they worked very well. 7. Which detection system did you use? Substrate? HRP, OPD from Sigma 8. Did you apply positive and negative controls along with the samples? Please specify. What did you use for standards? These are control tests. 9. Optimization attempts How many times have you tried the ELISA? Twice. Do you obtain the same results every time? Yes. What steps have you altered? No.

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Answer

Thank you for getting back to us and providing more details of your protocol. We are very sorry to hear that you are not happy with our product. We have searched our database and found that this is a popular selling product and your feedback is the first we have received about it not working. Therefore, at this stage, we would suggest that there is either a problem with the vial you received, or modifications to your protocol are needed to obtain a positive result. You have mentioned in your e-mail that you performed direct ELISA. For this type of ELISA, you need to coat the well with the antigen rather than with the primary antibody. For coating buffer, we would suggest using 0.1 M bicarbonate buffer (pH 9.2) instead of Tris buffer. It stabilizes coated proteins by maintaining their tertiary three-dimensional structure. By stabilizing the adsorbed protein, antigenic regions are preserved, allowing for greater binding reactivity with the detection molecule, thereby enhancing the specific signal. We hope you find this information useful.If you need anything further or any help then please let us know.

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Question

1. Please describe the problem (high background, no signal, non-specific colour development, poor standard curve etc). Signal is weaker than normal antibody. 2. What type of ELISA are you performing? (Direct ELISA, indirect ELISA, sandwich ELISA etc.) Direct ELISA. 3. On what material are you testing the antibody in ELISA? Species? Cell type? etc? Coat the plate with primary Antibodies. 3. How did you coat the plates? Coating buffer: 20mM Tris-HCL pH8.5 100ul/well, primary Ab 1:1000. 4. How did you block the unspecific binding sites? For how long? How did you wash the wells? 3% BSA in TBS, 4 hours, wash by TBST buffer 5X between each step. 5. Primary antibody Specification (in which species was it raised against)? rat anti-HA, mouse anti-BDG At what dilution(s) have you tested this antibody? 1:1000, 1:500 Incubation time, wash steps (multiple short washes are more effective than fewer longer wash steps)? 2 hours, 5X TBST 6. Secondary antibody What secondary antibody are you using? Your ab6120-250, ab5870-250. Specification (in which species was it raised against)?goat At what dilution(s) have you tested this antibody? Incubation, wash steps? 1 hour, 5XTBST Do you know whether the problems you are experiencing come from the secondary? I also used second antibody from Pierce as a control, they worked very well. 7. Which detection system did you use? Substrate? HRP, OPD from Sigma 8. Did you apply positive and negative controls along with the samples? Please specify. What did you use for standards? These are control tests. 9. Optimization attempts How many times have you tried the ELISA? Twice. Do you obtain the same results every time? Yes. What steps have you altered? No.

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Answer

Thank you for getting back to us and providing more details of your protocol. We are very sorry to hear that you are not happy with our product. We have searched our database and found that this is a popular selling product and your feedback is the first we have received about it not working. Therefore, at this stage, we would suggest that there is either a problem with the vial you received, or modifications to your protocol are needed to obtain a positive result. You have mentioned in your e-mail that you performed direct ELISA. For this type of ELISA, you need to coat the well with the antigen rather than with the primary antibody. For coating buffer, we would suggest using 0.1 M bicarbonate buffer (pH 9.2) instead of Tris buffer. It stabilizes coated proteins by maintaining their tertiary three-dimensional structure. By stabilizing the adsorbed protein, antigenic regions are preserved, allowing for greater binding reactivity with the detection molecule, thereby enhancing the specific signal. We hope you find this information useful.If you need anything further or any help then please let us know.

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