Goat F(ab')2 Anti-Mouse IgG - H&L (HRP), pre-adsorbed (ab5870)

Overview

  • Product name
    Goat F(ab')2 Anti-Mouse IgG - H&L (HRP), pre-adsorbed
    See all IgG secondary antibodies
  • Host species
    Goat
  • Target species
    Mouse
  • Tested applications
    Suitable for: ICC/IF, Dot blot, WB, ELISA, Electron Microscopy, IHCmore details
  • Minimal
    cross-reactivity

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

    Mouse IgG whole molecule.

  • 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

    BSA is IgG and protease free
  • Concentration information loading...
  • Purification notes
    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.
  • Conjugation notes
    Horseradish Peroxidase (HRP) (Molecular Weight 40,000 daltons)
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

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.

Application Abreviews Notes
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.

References

This product has been referenced in:
  • 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). Read more (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 . Read more (PubMed: 27207636) »
See all 3 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

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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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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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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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.

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

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