Goat Anti-Mouse IgG1 gamma chain (Cy5 ®) preadsorbed (ab136127)

Overview

  • Product name

    Goat Anti-Mouse IgG1 gamma chain (Cy5 ®) preadsorbed
    See all IgG1 secondary antibodies
  • Description

    Goat polyclonal Secondary Antibody to Mouse IgG1 - gamma chain (Cy5 ®), pre-adsorbed
  • Host species

    Goat
  • Target species

    Mouse
  • Tested applications

    Suitable for: ELISA, Flow Cyt, WB, ICC/IFmore details
  • Minimal
    cross-reactivity


    Human more details
  • Immunogen

    Mouse IgG1 paraproteins

  • Conjugation

    Cy5 ®. Ex: 650nm, Em: 667nm

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C.
  • Storage buffer

    Preservative: 0.1% Sodium azide
    Constituent: 99% PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    ab136127 was purified by affinity chromatography on mouse IgG1 covalently linked to agarose and cross-absorbed with Mouse IgM, IgG2a, IgG2b, IgG3 and IgA, pooled Human sera and purified Human paraproteins.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab136127 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
ELISA Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
WB Use at an assay dependent concentration.
ICC/IF Use 0.1µg for 106 cells.

References

This product has been referenced in:

  • Schmitt D  et al. Initial characterization of a Syap1 knock-out mouse and distribution of Syap1 in mouse brain and cultured motoneurons. Histochem Cell Biol 146:489-512 (2016). Read more (PubMed: 27344443) »
  • Dombert B  et al. Presynaptic localization of Smn and hnRNP R in axon terminals of embryonic and postnatal mouse motoneurons. PLoS One 9:e110846 (2014). Read more (PubMed: 25338097) »
See all 2 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

The concentration of this lot is GR91861-1 is 0.85mg/ml. An appropriate isotype control would be ab91353, Mouse IgG1, kappa monoclonal [MG1-45] - Isotype control. This product is shipped with a concentration of 1mg/ml. https://www.abcam.com/Mouse-IgG1-kappa-monoclonal-MG1-45-Isotype-control-ab18447.html We recommend use of a pre-absorbed secondary antibody in Flow Cytometry. We have a number of pre-absorbed anti-Mouse IgG1 antibodies which are conjugated to fluorescent molecules for use in Flow. Choice of the appropriate marker will depend upon your experimental set up and flow cytometer. I have complied a short list for you below. Secondary Antibodies: AB98692 (FITC conjugated): https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG1-heavy-chain-FITC-pre-adsorbed-ab98692.html AB136127 (Cy5 conjugated): https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG1-gamma-chain-Cy5--pre-adsorbed-ab136127.html AB130785 (AP/Cy7 conjugated): https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG1-heavy-chain-AllophycocyaninCy7-pre-adsorbed-ab130785.html AB130786 (AP conjugated): https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG1-heavy-chain-Allophycocyanin-pre-adsorbed-ab130786.html

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Question

Hi,



I am trying to perform a modified sandwich ELISA/FluoSpot assay.

Basically what I want to do is coat wells in 96 or 384 well plate with an antibody (a mouse IgE,k) against protein X.

Then I wish to grow cells expressing protein X in the well.

Then after they have expressed some protein X, I wish to add a second antibody (a mouse IgG1,k) against protein X.

And finally a fluorescently conjugated secondary antibody against the second antibody (an anti mouse IgG1).

Then I will look in a microscope and identify cells producing large quantities of protein X and using a special technique I will purify those single cells.

Everything will preferably happen in cell media, preferably with serum, but if that is not feasible PBS can be used for the actual detection part (second antibody, secondary antibody, purification).



I am considering your “Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Cy5 ®), pre-adsorbed (ab6563)”.

Is there any chance this would cross react with the first antibody (mouse IgE)?

Is there any reason to believe this antibody would not work in the experiment above?

Also, I have to use a specific vendor for plates. They only supply “plastic” bottom plates. In your ELISPOT protocol you mention PVDF or nitrocellulose bottom plates. What is the point of the PVDF/nitrocellulose bottoms? I know that protein G can adhere to The “plastic” bottom, so I would expect antibodies would adhere as well or have I misunderstood something?



Kind Regards,

Read More
Answer

Thank you for your inquiry.

I used to perform many ELISPOTs by hand as you with plastic plates. The capture antibody was coated on the plates. The PVDF membrane bottom well plates areused when using a reader to analyze the results. The membranes are pushed through onto sticky tape and read in a reader with special software to identify spots.

I am not aware of a technique where the cells can be identified and sorted according to the production amount of protein X. I am assuming that protein X is secreted and therefore should be captured in the vicinity of the producing cell, but any slight movement of the plate will swirl the cells and make a identification impossible.

ab6563 is in general suitable as secondary to a mouse IgG1. But is is anti heavy and light chain. So it will pick up the light chain too. I suggest to choose an antibody specific to the heavy chain. I can recommend ab136127:

https://www.abcam.com/index.html?datasheet=136127 (or use the following: https://www.abcam.com/index.html?datasheet=136127).

This antibody is not specifically cross absorbed against IgE, but against Mouse IgM, IgG2a, IgG2b, IgG3 and IgA, pooled Human sera and purified Human paraproteins. ab136127 is tested in ELISA and therefore should also work in a ELISPOT setting as secondary.

I hope this information is helpful. Please do not hesitate to contact me again with any further questions in this matter.

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