Biotin Rat IgG1, kappa [RTK2071] - isotype control (ab18403)

Q&A (2)

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FITC Unconjugated

Overview

  • Product name

    Biotin Rat IgG1, kappa [RTK2071] - isotype control
    See all Rat isotype controls

  • Conjugation

    Biotin

  • Conjugation notes

    The antibody was conjugated with biotin under optimal conditions. The solution is free of unconjugated biotin.

  • Specificity

    The RTK2071 immunoglobulin reacts with KLH. The RTK2071 immunoglobulin was chosen as an isotype control after screening on a variety of resting, activated, live, and fixed mouse and human tissues.

  • Tested applications

    Suitable for: Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Trinitrophenol and KLH.

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.

  • Storage buffer

    pH: 7.20
    Preservative: 0.09% Sodium azide
    Constituent: PBS
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    RTK2071

  • Isotype

    IgG1

  • Light chain type

    kappa

  • Research areas

  • Alternative names

    • Rat Isotype Control

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab18403 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
Notes
Application notes
Flow Cyt: Use at an assay dependent dilution for cell surface or intracellular staining.

Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.

References (0)

Publishing research using ab18403? Please let us know so that we can cite the reference in this datasheet.

ab18403 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Q&A
Submit a question

1-2 of 2 Q&A

Question

Thank you for your help.
I have two additional questions:
- what do you mean as " - no primary - only secondary control" ?
- can you provide a protocol to use the isotype control you suggest?
Thank you for your collaboration

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Answer

Thank you for your response.

"No primary control" means that you perform the staining protocol as normal (fixation, blocking, secondary antibody, detection) - except you need to omit the incubation step with the primary antibody. Instead of applying the primary antibody, you only incubate the samples with the dilution buffer you would normally use for diluting the primary antibody.

Isotype control meansyou perform the staining protocol as normal butt the samples will be treated with the isotype control instead of the primary antibody. The isotype of this primary antibody is IgG1, so the isotype control is rat IgG1 .

I would advise you to visit this site to get some further information about positive control and isotype control:

https://www.abcam.com/index.html?pageconfig=technicalfaqs

I hope this helps and if I can assist further, please do not hesitate to contact me.

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Question

Inquiry: Dear custumer service, we recently used your HCN4 antibody (ab32675) to perform an immunofluorescence assay on human atrial and ventricular myocytes. Staining was succesful and this data were submitted for publication. One of the reviewer asked to have a negative control, e.g. a cell stained with the HCN4 antibody pre-absorbed with the specific peptide. How can we meet this request? do you have a specific peptide and a suitable protocol to perform this assay? thank you for your assistance. Best

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Answer

Thank you for your enquiry and your interest in our products.

The immunogen used to raise this antibody was a synthetic peptide: SHGSLLLPPA SSPPPPQVPQ RRGTPPLTPG RLTQDLKL, corresponding to amino acids 1048-1085 of Human HCN4:SHGSLLLPPASSPPPPQVPQRRGTPPLTPGRLTQDLKL. Unfortunately, the peptide is not commercially available.

Even though the immunogen peptide is not available, you could apply several controls to check the specificity of this antibody:

- isotype control,

- negative control,

- no primary - only secondary control

The isotype of this antibody is IgG1 so you can apply an isotype control using rat IgG1 (ab46745, ab128477, ab18403, ab18404, ab18407).

HCN4 is highly expressed in thalamus, testis and in heart, both in ventricle and atrium. It is detected at much lower levels in amygdala, substantia nigra, cerebellum and hippocampus. Generally, negative controls are cells/tissue which do not express the target protein or at very low level. You may find some useful information about expression level of HCN4 at this site: http://www.proteinatlas.org/ENSG00000138622

I hope this helps and if I can assist further, please do not hesitate to contact me.

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