Anti-Interferon beta antibody (Biotin) (ab84258)

Rabbit polyclonal Interferon beta antibody conjugated to Biotin. Validated in WB, IHC, ICC/IF and tested in Human. Cited in 2 publication(s).

Overview

  • Product name
    Anti-Interferon beta antibody (Biotin)
    See all Interferon beta primary antibodies
  • Description
    Rabbit polyclonal to Interferon beta (Biotin)
  • Host species
    Rabbit
  • Conjugation
    Biotin
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Highly pure (>98%) recombinant human Interferon beta

  • Positive control
    • This antibody gave a positive signal in human liver, spleen and kidney tissue lysates.

Properties

Applications

Our Abpromise guarantee covers the use of ab84258 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 27 kDa (predicted molecular weight: 22 kDa).
IHC-P Use a concentration of 0.75 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • Function
    Has antiviral, antibacterial and anticancer activities.
  • Sequence similarities
    Belongs to the alpha/beta interferon family.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • beta-interferon antibody
    • Fibroblast interferon antibody
    • IFB antibody
    • IFF antibody
    • IFN beta antibody
    • IFN-beta antibody
    • IFNB 1 antibody
    • IFNB antibody
    • IFNB_HUMAN antibody
    • IFNB1 antibody
    • Interferon beta 1 fibroblast antibody
    • Interferon beta antibody
    • Interferon beta precursor antibody
    • MGC96956 antibody
    see all

Images

  • All lanes : Anti-Interferon beta antibody (Biotin) (ab84258) at 1 µg/ml

    Lane 1 : Human liver tissue lysate - total protein (ab29889)
    Lane 2 : Human spleen tissue lysate - total protein (ab29699)
    Lane 3 : Human kidney tissue lysate - total protein (ab30203)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 22 kDa
    Observed band size: 27 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 100 kDa, 46 kDa, 55 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 16 minutes


    The Interferon Beta protein sequence contains both a phosphorylation and a glycosylation site (Swissprot). Post-translation modifications could account for the migration of the band at a higher molecular weight than predicted.
  • ICC/IF image of ab84258 stained HepG2 cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab84258, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human normal placenta, staining Interferon beta with ab84258 at 0.75 µg/ml. An HRP-labeled polymer detection system was used with a DAB chromogen. Heat induced antigen retrieval with a pH 6.0 sodium citrate buffer is recommended.

References

This product has been referenced in:
  • Liu Y  et al. Suppression of mRNA Nanoparticle Transfection in Human Fibroblasts by Selected Interferon Inhibiting Small Molecule Compounds. Biomolecules 7:N/A (2017). Read more (PubMed: 28758979) »
  • Price JV  et al. Protein microarray analysis reveals BAFF-binding autoantibodies in systemic lupus erythematosus. J Clin Invest 123:5135-45 (2013). Read more (PubMed: 24270423) »
See all 2 Publications for this product

Customer reviews and Q&As

1-5 of 5 Q&A

Answer

Thank you for your reply.

I do understand your concerns. It may be that you would want to run a western blot with this product to ensure that it is picking up IFN from your cells, it would them be possible to run a purified protein as control since we are unsure the hepatocytes would express. If this worked you could then stain with more confidence.

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!

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Answer

Thank you very much for your interest in ab84258.

Unfortunately the sequence homology between he human and mouse forms of IFNB is very low (47%). We would not be confident that this product would work in mouse.

We do have another product ab24324 (https://www.abcam.com/Interferon-beta-antibody-7F-D3-ab24324.html) which is tested and guaranteed to work in mouse. However we have not yet tested this in IF. I can offer a discount off a future purchase if you buy ab34324 now, test it in IF and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free PRIMARY ANTIBODY.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab24324 in IF. I will then send a discount code. This code must be issued before purchasing ab24324 so please wait for my reply before ordering.

2. Purchase ab24324 either by phone, fax, or online (www.abcam.com).

3. Test it in IF.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any PRIMARY ANTIBODY ordered and the discount code is valid for 4 months after issue.

Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab24324 turns out to be unsuitable for IF, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.


The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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Answer

Thank you for contacting us.

I think these two antibodies will work together in a sandwich ELISA, but we have not tested them together. You will need to test a few different dilutions to achieve the optimal signal with the least background.

I suggest testing combinations of capture antibody ab6979 coated onto the plate at concentrations of 0.5, 0.1, and 0.02 ug/ml (1/2000, 1/10,000, 1/50,000 of the 1mg/ml stock concentration).

and detection antibody ab84258 applied to captured samples at concentrations of 0.04, 0.2, and 1.0 ug/ml (Dilutions will be dependent on the stock concentration of the reconstituted antibody).

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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Question
Answer

Thank you for contacting Abcam. I have heard back from the lab and they have told me that the biotin is covalently coupled to primary amines (lysines) of the immunoglobulin and not to either the N or C terminal. If there is anything else I can help you with, please let me know.  

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