Creative Biolabs has amassed considerable expertise in the screening of IHC-positive hybridomas.

Introduction of IHC-Positive Hybridoma Screening Services

Immunohistochemistry (IHC) represents a pivotal antibody-based methodology employed to ascertain protein expression within tissues. Its ubiquitous application spans both medical research laboratories and clinical environments, facilitating the interrogation of antigen presence within biological specimens. IHC hinges upon the precise detection of antibodies directed against specific epitopes, thereby imposing heightened requisites on antibody generation. Within our hybridoma screening framework, Creative Biolabs meticulously cultivates IHC-specific antibodies through dedicated IHC-positive hybridoma screening. Leveraging a sophisticated and exhaustive hybridoma platform, we extend a spectrum of comprehensive hybridoma experimental services tailored to your needs.

In our pursuit of antibodies suitable for IHC or immunocytochemical staining (ICC), beyond employing native immunogens like recombinant proteins, we also explore the utilization of denatured antigens during animal immunization and hybridoma screening. Our hypothesis revolves around the notion that antigens targeted for IHC staining undergo denaturation, either wholly or partially, along with alterations induced by cross-linking or precipitation during paraffin embedding and various fixation procedures. Consequently, we may opt for fixative-treated immunogens during animal immunization. Ultimately, we conduct hybridoma screening tailored for IHC, utilizing paraffin-embedded slides or tissue microarrays. Clients have the option to provide their own slides or tissue arrays or use the slides or arrays in Creative Biolabs.

Approaches of IHC-Positive Hybridoma Screening

In the realm of IHC, elucidating the specificity of antibodies poses a formidable challenge, primarily due to the scarcity of ideal controls. Wherein, gene-knockout tissues or specific transgenic fusion proteins serve as viable references for staining. Consequently, an ancillary assay, such as ELISA, Western Blotting (WB), or IP, becomes imperative to corroborate the antibody's binding specificity to the target antigen. Subsequently, the antibody undergoes scrutiny in IHC assays, demanding specificity in staining—manifested by positivity in select tissues or cells, with staining intensity surpassing that of isotype-matched negative control antibodies. To obtain antibodies suitable for IHC, it is essential to conduct an initial screening using ELISA, Western blotting (for epitopes in denatured conditions), and/or immunoprecipitation (for epitopes in native conditions). Empirical evidence suggests a high likelihood of obtaining IHC-positive clones when multiple ELISA/WB/IP-positive clones are secured.

Features of IHC-Positive Hybridoma Screening in Creative Biolabs

Creative Biolabs delivers antibodies of exceptional specificity:

• Elevated signal-to-noise ratios ensure optimal performance.
• Staining patterns accurately reflect the confirmed localization of the target protein in control cells or tissues.
• Immunoblots reveal a solitary band, indicative of precise antigen recognition.
• Discovery of novel epitopes.
• Exceptional results, even with complex antigens.
• Access to a comprehensive range of assays, encompassing ELISA, FACS, ICC, IHC, WB, and flow cytometry.

FAQ

1. Q1. How does Creative Biolabs screen hybridomas for IHC positivity?

   A: Creative Biolabs employs a rigorous screening process to identify IHC-positive antibodies. Initially, antibodies undergo assessment via ELISA, Western Blotting, and/or immunoprecipitation to identify those demonstrating favorable binding properties. This preliminary selection helps increase the likelihood of identifying antibodies suitable for IHC assays.

2. Q2. What challenges are associated with determining antibody specificity in IHC assays?

   A: Determining antibody specificity in IHC can be challenging due to the lack of perfect controls. While a gene-knockout tissue or a specific transgenic fusion protein often serves as a benchmark for staining control, independent assays are necessary to verify the antibody's specific binding to the target. Antibodies that pass these assays must demonstrate specific staining in IHC, showing positivity in selected tissues or cells and significantly stronger staining compared to isotype-matched negative control antibodies.

3. Q3. What are the final deliverables of Creative Biolabs' IHC-Positive Hybridoma Screening services?

   A: Our service will eventually supply the customer with 1-3 positive cell lines, 1-3 mg of antibody, the ELISA titer exceeded 1:50,000, and the Western Blot test was positive, a detailed report, and the residual antigens.

Endowed with extensive proficiency in mAb production, Creative Biolabs emerges as a venerable leader in this field. Supported by our state-of-the-art hybridoma facilities, we offer a comprehensive suite of hybridoma-based services. Should our offerings pique your interest, we wholeheartedly invite you to contact us for additional clarification.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.