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Guarantee Statement

Bethyl's purpose is to improve lives by supporting scientific discovery. This drives our complete dedication to the highest standards of product and service quality.

Bethyl is unique in our end-to-end control of the entire antibody lifecycle. From our veterinary facilities to the development, production and validation labs, the Bethyl team focuses on delivering quality products. This attention to quality at every step and the integration across each stage of the process means scientists can rely on Bethyl antibodies. We also offer expert technical support to help ensure your success.

Recognizing that your research and time are valuable, we promise to stand behind our products. Should one of our antibodies fail to return satisfactory results when used in the species, application and under the same conditions stated on the product datasheet, Bethyl will offer a full product replacement, credit or refund at the investigator's discretion. Before providing product replacement, credit or refund, we ask the investigator to complete an application specific questionnaire and work with our Technical Support team to identify potential steps to optimize conditions.

Every Bethyl product ships with a 100% guarantee; choose Bethyl antibodies with confidence.

Product Validation

  1. Affinity and Specificity
    1. Methods for the purification of antibodies include: physiochemical fractionation, class-specific affinity chromatography and antigen-specific affinity chromatography. All primary antibodies produced at Bethyl are raised against a peptide and purified via antigen-specific affinity chromatography. To yield antibodies of high affinity and specificity, Bethyl has developed proprietary methods and buffers for many aspects of the affinity purification process. In this process, the peptide used for immunization is immobilized on agarose beads. Serum is passed over a column of these antigen-specific beads, and antibodies with a high affinity to the peptide are captured. All other serum components and non-specific antibodies are washed away, leaving a high-affinity product. Antibodies are then eluted in a purified form, ensuring products have a high affinity to the target protein.
    2. Bethyl manufactures highly specific products due to a clearly defined and refined immunization schedule. Boosting the host over many weeks encourages the immune system to continually polish B-cell antibody production, progressively enhancing antibody specificity.

       

  2. Validation
    1. The first and primary goal for antibody validation is confirming that the antibody recognizes its intended protein target. To qualify antibodies for market, Bethyl relies on a strategy that involves reciprocal immunoprecipitation (IP) and Western blot (WB). When generating peptide-based antibodies, two to four peptides from different regions of the same target protein are used to generate individually affinity purified antibodies.

      Reciprocal IP and WB entails the immunoprecipitation of the target protein by each of the polyclonal antibodies generated against the specific target. The immunoprecipitates are then immunoblotted with each of the generated antibodies. Validation is confirmed via the results of the reciprocal IP and WB experiments. Antibodies directed against disparate regions of a target should exhibit recognition of a common “band” developed on the immunoblot. Relative mobility of target band(s) is used as a secondary confirmation that the antibody is recognizing its intended target.

      Immunoprecipitation and detection of endogenous protein from appropriate cell lysates is always performed. Detection of purified recombinant protein or target protein in overexpressing lysates is not performed as this may result in a false sense of success, quality and utility of the antibody.

    2. Because Bethyl uses IP/WB as the benchmark for product release, there existed a desperate need to find a solution to the problems presented by the IgG noise that is prominent in traditional methods for WB of immunoprecipitates. To overcome this limitation, the ReliaBLOT® Reagents and Procedures were developed. Currently, the ReliaBLOT® Reagents and Procedures are incorporated as the default protocol for IP/WB performed in Bethyl’s testing lab.
       
  3. Lot-to-Lot Consistency
    1. Side-by-side comparison by IP/WB assays of previous lots and new lots are performed to assess lot-to-lot consistency. Each new lot must perform similarly to the previous lots. If the new lot does not perform similarly, it will not be released to the market.

      Demonstration of Lot-to-Lot Consistency by Immunoprecipitation/Western Blot (IP/WB)

      Demonstration of Lot-to-Lot Consistency by Immunoprecipitation/Western Blot (IP/WB).
      A300-033A lot 4, A300-033A lot 5, and normal rabbit IgG were used in immunoprecipitations (IPs) of HeLa whole cell lysate (WCL). A portion of the IPs along with 50 µg HeLa WCL were blotted and incubated in either lot 4 or lot 5 antibody at equal concentrations (1µg/ml). Exposure times to a CCD camera were also equal.