Mouse IgG1 monoclonal (Phycoerythrin) - Isotype control  (ab81200)

Overview

  • Product name

    Mouse IgG1 monoclonal (Phycoerythrin) - Isotype control 
    See all Mouse isotype controls
  • Conjugation

    Phycoerythrin. Ex: 488nm, Em: 575nm
  • Conjugation notes

    R-PE exitation wavelength 488 nm, emission wavelength 575 nm is the correct sentence.
  • Tested applications

    Suitable for: Flow Cytmore details
  • General notes

    Reagents should be brought to room temperature (22 ± 3°C) before use.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
  • Storage buffer

    Preservative: 0.1% Sodium azide
    Constituents: 0.4% BSA, 1.74% Sodium chloride, 0.328% Sodium phosphate
  • Concentration information loading...
  • Purification notes

    Purified from mouse myeloma useing 0,22 µm filtration
  • Clonality

    Monoclonal
  • Isotype

    IgG1
  • Research areas

  • Alternative names

    • Mouse Isotype Control

Applications

Our Abpromise guarantee covers the use of ab81200 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
Flow Cyt Use at an assay dependent concentration.

Nonspecific binding in flow cytometric methods of various diseases.

References

This product has been referenced in:

  • Bauer K  et al. Slit-2 facilitates interaction of P-cadherin with Robo-3 and inhibits cell migration in an oral squamous cell carcinoma cell line. Carcinogenesis 32:935-43 (2011). Read more (PubMed: 21459757) »
See 1 Publication for this product

Customer reviews and Q&As

1-4 of 4 Q&A

Answer

Thank you for contacting Abcam.

Please find attached the Certificate of Compliances that were requested.

If there is anything else I can help you with, please let me know.

Read More

Answer

Thank you for contacting us.

As requested, I have included the Certificate of Compliance for each of the products listed.

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

Read More

Answer

Thank you for contacting us. If you are not able to easily pellet the exosomes, I would indeed recommend to add 20 µl of the antibody solution to the exosomes in 100 µl PBS. You may need to increase the incubation time and or the volume of ab81436 added, in order to optimise the staining conditions for this particular setting. To our knowledge, this has never been done before, so I am sorry that we cannot give you more detailed information. However, I trust that you will succeed! Unfortunately, it is NOT possible to determine the EXACT concentration of ab81436: The concentration of the specific TAPA1 antibodies has not been determined before conjugation. Even if it had been, this concentration would be different from the concentration of the antibody after the conjugation procedure, as there is always a loss during the purification steps. Determining the protein concentration after the conjugation does not result in accurate measurements, because Phycoerythrin (PE) is a huge protein itself and thus would give false results, as my colleagues tried to explain earlier (in case you did not receive these emails, see below). We would therefore suggest to use the PE isotype control ab81200 in the same dilution than ab81436. For example, if you add 20 µl of ab81436 to 100 µl of your sample, than it is recommended to also use 20 µl of the isotype control ab81200 for 100 µl of sample. Also, I would like to draw your attention to the criteria to record information about the experimental overview and samples (including controls) set out by the flow cytometry community (e.g. http://www.flowcytometryuk.org/). These criteria are known as the Minimum Information about a Flow Cytometry Experiment (MIFlowCyt), and contain valuable information about isotype controls and FMO controls. I hope this information is helpful to you. Please do not hesitate to contact me if you need any further advice or information in this regard. Previously: I can confirm that the concentration of this antibody was not determined. Regrettably, it is not possible to correctly measure the antibody concentration of products with antibodies conjugated to proteins (such as PE). /If there are concentrations given for conjugated antibodies, it is usually the IgG concentration measured before the conjugation took place and therefore only an estimate./ Unfortunately, we do not currently have more information about the concentration of ab81436./ Antibody concentration is usually determined by measuring the protein concentration. This is not useful in this case since PE is also a protein.

Read More

Answer

Thank you for contacting us. I can confirm that the PE-conjugated CD81 antibody ab81436 is designed for Flow Cytometry analysis of human blood cells, and if the volume of your whole blood sample is 100 µl as indicated, we would suggest the following procedure: centrifuge the 100 µl of whole blood, remove the supernatant and keep the pellet optional step: perform a red blood lysis (see attached for details) add 20 µl of the ready-to-use antibody incubate and wash as usual and proceed with analysis Also, if you perform you perform flow cytometric analysis, we would recommend to optimise the instrument settings in order to detect the correct particles, as exosomes can be much smaller than cells. For your questions about the isotype controls: Isotype controls are a type of control designed to measure the level of non-specific background signal caused by primary antibodies. The background signal is usually the result of immunoglobulins binding non-specifically to Fc receptors present on the cell surface. For example, antibodies raised in mice, particularly those of the IgG2a isotype, will bind strongly to some human leukocytes regardless of the test antibody specificity. In this case you would need a mouse IgG2a isotype control for use with human cells or tissues. Thus, the isotype control antibody should always match the primary antibody’s host species, isotype, AND conjugation. It is therefore not advisable to use a FITC-conjugated IgG1 isotype control for a PE-conjugated antibody. Usually, if the concentration of the specific antibody is unknown, it can be recommended to use the isotype control in excess, i.e. to use a lower dilution for the isotype control than for the antibody of interest. Although isotype controls are frequently used, it is not the only type of control to determine non-specific binding and positivity for the marker of interest. This control is termed fluorescence minus one or FMO and works like an isotype control without the actual antibody. FMO controls are set up by leaving out one of the antibodies in your staining panel. This control might be also an option for your experiments and you may find more information about this type of control in the latest publications or on flow cytometry websites, such as http://www.isac-net.org or http://www.cyto.purdue.edu/ . I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up