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Technical support

Frequently asked questions

Production

Delivery and availability

Quality Control

Protocols

  • How do we dissolve the immunizing peptide and how do we mix it with the antibody?
  • There is a UN setting (UN Number) for Sodium Azide. Should we follow the setting as it is less than 0.1%? Would it be hazardous?
  • How do I store my antibody?
       
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    What is the isotype of the Everest antibodies? 

    All our goat antibodies are polyclonal antibodies and they are affinity purified using the immunizing peptide. Consequently the end product will be primarily IgG. When one wants to bind the antibody to a stationary phase, we recommend protein G.


    What is the expected yield per goat? 

    Yield strongly depends on the target protein, the individual goat, the choice of peptide and environmental factors. Yield can be as low as a few mg and as high as a few hundreds of mg. We never exceeded 400 mg levels from one single goat. Please bear in mind that we are dealing with peptide affinity purified IgG developed to a 10-15aa peptide, so our yields cannot be fairly compared with those developed to total proteins, or with protein A/G purified antibodies. For high yield demand, please view our custom antibody  services page.


    Will you be able to provide grams of affinity purified IgG? 

    See above. For grams we will need to immunize several animals and pool the yield. It would make sense to test a sample from an existing product from our catalogue before ordering grams of the same. Please bear in mind that each animal may produce a slightly different product. By pooling the products from several animals (all immunized with the same peptide) we would dilute any individual variations and the blend will be the gold standard for future productions. For more details on our custom services, please view our custom antibody services page.


    What is the formulation of the antibodies? 

    All our un-conjugated antibodies are delivered as 100µg of purified antibody in 200µl buffer: 0.5 mg IgG/ml in Tris saline (20mM Tris pH7.3, 150mM NaCl), 0.02% sodium azide, with 0.5% bovine serum albumin.  If you would like to place a bulk order (1mg or over) with a different formulation (eg for applications where BSA would be in the way)  please contact sales@everestbiotech.com.


    Can we obtain the immunizing peptide? 

    Yes, we have many peptides in stock, ready for next day delivery. Related peptides can be found on each antibody product page. If you cannot find what you're looking for please contact sales@everestbiotech.com.


    Can I get a free antibody sample to try? 

     

    In most cases we do not have trial size samples available. When we do, the purchase of such sample will be attractively priced. Both the full size product and the trial antibody are guaranteed, so you get your money back when the antibody fails to meet your expectations (see 100% guarantee).

    For terms and conditions please contact support@everestbiotech.com

     


    Does Everest provide custom service? 

    Yes, we can make you any antibody you need with exclusivity. We will immunize two goats for each immunogen for this service, but any deviation can be negotiated (more info here).


     

    Do you deliver antibodies in bulk? 

    Yes, and we can deliver BSA-free (more info here). This is particularly interesting for assay developers, as a bulk order will prevent batch-to-batch variations during the project. Bulk orders are accepted from 1mg and higher. When grams quantities are required, a regular custom project involving immunization of larger animals will have to be involved. Have a look at our custom antibody services page.


    What secondary antibody is recommended? 

    We recommend affinity purified IgG with minimal cross reactivity to human and rodent serum proteins. We sell the secondary antibodies that we use in our QC department (see http://everestbiotech.com/category/secondary-antibodies/).


    Can I get a discount? 

    We offer 20% discount on your first order if your sign up to our newsletter. If you wish to buy in bulk, please view our pricing here.


    Do you have distributor in my country? 

    To view a distributor in your country, please select your country in top right corner of the page and then click on Contact us. If you can't find your country on the list, please email sales@everestbiotech.com.


    Will you be able to provide grams of affinity purified IgG? 

    See above. For grams we will need to immunize several animals and pool the yield. It would make sense to test a sample from an existing product from our catalogue before ordering grams of the same. Please bear in mind that each animal may produce a slightly different product. By pooling the products from several animals (all immunized with the same peptide) we would dilute any individual variations and the blend will be the gold standard for future productions. For more details on our custom services, please view our custom antibody services page.


    Can we get pre-immune serum? 

    No. There are two reasons: We cannot export serum or plasma from the country of origin. Since the product is affinity purified using the immunizing peptide, testing pre-immune serum is not relevant. When the product is any good, any non-specific background need to be addressed by omitting the primary in order to establish the background generated by the secondary. Once this is done, reducing primary incubations times (never incubate overnight), diluting the primary further and optimizing the blocking conditions (always incubate primary in the blocking buffer) need to be tested. When non-specific background persists contact support@everestbiotech.com. The unexpected presence of one or two extra bands may not be non-specific background. We can provide immunizing peptide to block these signals.


    Does the antibody cross-react to the species of my project? 

    All our antibodies are generated and affinity purified using a 10-15aa peptide. The amino acid sequence of this peptide determines whether the antibody cross-reacts across different species. A quick and easy assessment is to blast the sequence of the immunizing peptide in NCBI. We do so at the stage of peptide design and we state the expected species cross-reactivity on the datasheet based on these blast results. However, we limit the statement of expected mammal species to Human, Mouse, Rat, Dog, Pig and Cow. Please note that we have a separate statement on the datasheet for TESTED species.


    Why does the detected band in Western blot not correspond to the calculated molecular weight? 

    All scientists should check for post-translational modifications in the protein of interest. Apart from the modifications of individual residues in the protein, potential cleavage of the protein into two or more fragments by proteases should be considered. And to make things even more complex, such fragments may possess modified residues. Therefore, an aberrant migration does not at all necessarily indicate non-specificity of the used antibody. The specificity of the antibody needs to be assessed by comparing two identical blots, one with the primary and one without the primary. Any signal derived from the secondary antibody will occur in both blots. When secondary-derived background is excluded, comparison between endogenous and knocked-down/knocked-out/ over-expressed levels should give reliable hints about the specificity of the aberrant band(s). Alternatively the antibody could be blocked with the immunizing peptide (see above).


    Why can I not reproduce the data as shown on the product sheet? 

    The Quality Control data as presented on the product sheet are guaranteed. When a customer cannot reproduce these data, please contact support@everestbiotech.com.
    When proper handling by the end user has been demonstrated, we may send a replacement, or we will arrange a refund.
    Different tissue types or cell lines may result in different patterns. Please try to copy the data as shown on the product sheet in order to confirm the integrity of the antibody. Please consult our protocols (see elsewhere in this section).


    Why did my last purchased vial not perform as good as previously purchased vials? 

    Please consider the previous question and answer. If this does not apply, then here is another explanation:
    Particularly when in high demand, a product sometimes reaches the end of its stock and needs replenishment by antibodies from a new goat. Despite the relatively short length of the immunizing peptide (10-15aa), each individual animal potentially generates a slightly different antibody and it needs to be quality assessed as if it is a completely new product. Required dilutions will differ, and sometimes the next animal will not generate identical QC results to the previous animal, although we strive to generate equivalent results for the product sheet. Hence we distinguish all our antibodies in batch codes. Each batch may represent a different round of peptide synthesis, a different conjugation event to the carrier protein, a different purification, or a different goat, or any combination of the above. We also distinguish different events of making aliquots from the same batch code by aliquot coding. Therefore, each vial has next to its catalogue EB-code a batch code and an aliquot code. Please specify these codes when you have a complaint to send to support@everestbiotech.com.


    The antibody shows high non-specific background. How do I reduce this background? 

    Please be aware that our antibodies are affinity purified and therefore needs slightly different handling from antiserum. Most scientists incubate their blots overnight with primary. Our products often generate non-specific background when used overnight, unless far further diluted. We always test our antibodies by labelling for one hour at room temperature. When the reactivity is too strong, dilute further in steps of a factor three. When ELISA titres to the immunizing peptide are up to 1:32000 usually starting concentration should be 1ug/ml for 1h incubations. Higher titres require higher dilutions and so would overnight incubations. See also our discussion on  non-specific background.


    I cannot find the Everest antibody in the paper cited on the product sheet. 

    We manufacture all the primary antibodies listed on this site but many are also sold through other vendors under their brands.   Where we have cited a paper that used our antibody under a different brand we prefix it with "The goat polyclonal antibody used in this paper was manufactured by us"


    I have a specific question about a product. What is the best way to contact you? 

    We prefer you contact us by email to support@everestbiotech.com. We will reply within 24 hours or less, Monday-Friday.


    Why do you only test ELISA titres and not specificity? 

    The ELISA titre is merely a measure to test the reactivity of the product to the immunizing peptide. It does not reflect its reactivity to the real target protein, nor does it measure specificity. These need to be addressed in the applications the antibody is required for. Both titre and specificity will be different for each application. Our titres are specified on the product sheets as: Peptide ELISA: antibody detection limit dilution. 


    For blocking experiments, how do we dissolve the immunizing peptide and how do we mix it with the antibody? 

    The antibodies come at 0.5mg/ml and peptide as a 100ug pellet. When peptides are reconstituted in 200ul water, the concentration would be also 0.5mg/ml. To start, the best ratio would be 1:1 (which means molar excess of peptides relative to antibodies when identical volumes are mixed).  See the procedure in PDF.


    There is a UN setting (UN Number) for Sodium Azide. Should we follow the setting as it is less than 0.1%? Would it be hazardous? 

    The tiny amount of sodium azide (NaN3) present in our products ensures that they can be transported under the "Excepted Quantity" exemptions from dangerous goods regulations. See the MSDS.


    How do I store my antibody? 

    Aliquot and store at -20°C. Minimize freezing and thawing. One working aliquot can be stored at 4C for regular use. The sodium azide will preserve the antibodies for 6-12 months as long as it is kept  cold and away from direct (sun)light.