Overview

  • Product name
    Goat Anti-Mouse IgG H&L (20nm Gold) preadsorbed
    See all IgG secondary antibodies
  • Host species
    Goat
  • Target species
    Mouse
  • Tested applications
    Suitable for: Electron Microscopymore details
  • Minimal
    cross-reactivity

    Cow, Horse, Human more details
  • Immunogen

    Full length Mouse IgG (H&L)

  • Conjugation
    Gold 20nm

Properties

Applications

Our Abpromise guarantee covers the use of ab27242 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
Electron Microscopy Use at an assay dependent dilution.

References

This product has been referenced in:
See all 7 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Thank you for contacting us.

These antibodies are raised against targets that are not species specific. The antibodies would detect the proteins irrespective of species so the Abtrial discount will not be valid.

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

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Answer

Gracias por contactarnos.

La dilución óptima a utilizar varía para cada caso en función de la muestra y la aplicación. Te sugiero usar un volumen pequeño tanto de muestra como de producto y probar un rango amplio de diluciones del secundario para determinar cuál de ellas os da la mejor señal sin causar un excesivo ruido de fondo. Esta será la dilución elegida para testar en tu aplicación.

El precio, como bien indicas es de 285€ más 22€ de gastos de envío.

La promoción que tenemos en la actualidad consiste en que al comprar cualquier anticuerpo primario os regalamos otro primario Rabbit Monoclonal. Simplemente indica el código RABMAB-XBSMG en el próximo pedido. Para más información picha https://www.abcam.com/index.html?pageconfig=resource&rid=15447.

Si tuvieras alguna duda o necesitaras ayuda para encontrar un RabMab apropiado, no dudes en contactarnos de nuevo.

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Answer

Thank you very much for contacting us with your questions, and also for your patience while I have been in touch with the lab regarding your enquiry.

For thegoat anti-mouse 20 nm conjugate ab27242, there areapproximately 48 IgG proteins attached to each particle. For thegoat anti-rabbit 40 nm conjugate,there areapproximately 192 IgG proteins per particle. Particle size distribution is measured via the coefficient of variance (CV%) statistic. For these products,we requirethe CV to be8% or less, so the particles are very monodisperse.

I hope that this information will be useful, but please let me know if you have any further questions and I'll be happy to help.

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Question

Re: Problems with Gold-conjugated secondary antibodies ab27241 and ab27242 in EM with neuroblastoma cells.

Hi, thanks for getting back to me.

The cells I use are neuroblastoma cells with rat origin. I transfect my cells with the GFP mutant synaptopHluorin, this enables me to image the hydrated cells by fluorescence microscopy.

I then fix my cells with PFA and permeabilised with Triton (I have tried concentrations between 0.1- 1%)

I then labelwith anti GFP primary antibody andgold conjugated secondary antibody to allow detection of the labelled areas in the hydrated cells by atmospheric SEM.

Generally the results show single particles of gold scattered throughout the cell. Even in samples where I have attempted to label actin filaments of endogenous proteins I do not see localisation to specific cellular patterns. It has also been questioned that surely more gold particles should be observed, taking into account the abundance of cellular structures labelled by the primary antibody. (Fluorescence assay have been carried out to determine the specificity of the primary antibody and have shown perfect colocalisation.)

I have seen better results when I permeabilse with 1% Triton, although here I used a new transfection agent, so far I have not been able to repeat these results.

I think a problem may be in either the specificity of the gold conjugated secondary antibody or the penetration of the gold particles into the hydrate cells. To solve this I can see two options 1) use smaller gold particles. This would be ok in normal SEMs although the resolution of my current instrument for ASEM is ˜20nm, so I'm not too keen to use much smaller particles.

2) Alternative methods in fixation/permeabilisation to allow easier passage.

I would be very grateful for your thoughts on my problem.

Thanks,

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Answer

Thank you very much for sending all these details which enable us to investigate the problem.

Thank you also for your patience while I was inquiring with the laboratory.

Firstly I can confirm that we have not had any otherreports of any problems with these towbatches ofantibodies mentioned.

In regards to solve the problem, I would like firstly to ask you whether you have checked whether the primary antibody is working. Have you used this primary antibody already on similar fixed slides and obtained a good signal?

In regards to two secondary antibodies, the laboratory recommends toreturnto using 1% Triton to rule out the change of transfection agent as being the cause of the issues.

We can also recommend(on advice of the laboratory)to try using a smaller gold particle (as for exampleab27244) and silver enhance this to obtain a larger signal which should be visible under the ASEM that they are using. The product needed to carry out this silver enhancing step is commercially available. If you would like us to source it for you, please let me know and I see what I can do. The silver enhancing step consists of simply adding an equal volume of the supplied silver enhancer and initiator to the sample after incubation with the secondary antibody (the conjugate being used) has been carried out. The silver particles will then attach to the gold particles which are present in the sample, growing them to a larger size and therefore making them large enough to be visible under ASEM.

In case that the suggestions will not improve the results, and the primary antibody is excluded as possible source of problem, we will be happy to replace or refund these antibodies for you in accordance with our Abpromise policy.

Please do let me know if you have any questions or concerns.

I am looking forward to hear back from you to know whether the problem has been resolved. Thank you for your cooperation.

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Answer

Thank you for your inquiry. Unfortunately, we do not currently have more information about this product. We will update the on-line datasheet for this product as soon as we receive additional data. We apologize for any inconvenience this may cause.

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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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