Rabbit recombinant monoclonal antibodies are identical immunoglobulins directed against a specific epitope. These antibodies provide a consistent alternative to polyclonal antibodies and traditional hybridoma monoclonals, with the added benefit of an in vitro development option. Invitrogen recombinant rabbit monoclonal antibodies are manufactured by transfecting mammalian cells with heavy and light chain antibody-coding cDNA. This highly reproducible process results in excellent lot-to-lot consistency, helping ensure optimal specificity and performance. 

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Why choose Invitrogen recombinant rabbit monoclonal antibodies?

With a wide range of antibodies available, we offer solutions for various applications, including western blotting, immunocytochemistry, flow cytometry, and more. Our antibodies undergo rigorous advanced verification processes to help ensure optimal performance, specificity, and sensitivity. This comprehensive validation helps provide researchers with confidence in the reliability and accuracy of our antibodies. By choosing Invitrogen recombinant antibodies, customers can access a diverse portfolio and trust in the quality and performance of our extensively verified products.

Recombinant rabbit monoclonal antibody production

Recombinant rabbit monoclonal antibodies are developed using molecular cloning and pairing of specific light and heavy chain genes to yield unique clones. The development process involves immunization of rabbits, followed by isolation and screening of peripheral blood mononuclear cells (PBMCs). The DNA from these cells is then cloned as a library and further screened using desired end-application. The heavy and light chain cDNAs from the functional positive clones are co-expressed using a mammalian expression cassette that has been optimized for efficiency. This resulting DNA is used to produce controlled in vitro derived antibodies through cell culture methods. (Figure 1).

Figure 1. Recombinant rabbit monoclonal antibody production.

Advantages of recombinant rabbit monoclonal antibodies

Recombinant rabbit monoclonal antibodies are produced using recombinant DNA technology, allowing for precise control and modification of the antibody sequence, while conventional rabbit monoclonal antibodies are generated through traditional hybridoma technology. Recombinant antibodies offer exceptional lot-to-lot consistency as they are derived from banked DNA, eliminating cell drift, and ensuring consistent expression patterns. They also demonstrate enhanced specificity and sensitivity due to advanced screening processes, while conventional antibodies may vary in these aspects. Furthermore, recombinant antibodies can be customized for desired characteristics, whereas conventional antibodies have limited customization options.

Better specificity and sensitivity compared to standard antibodies.

Invitrogen antibodies are subjected to a thorough and comprehensive advanced verification process, which involves the utilization of various methods such as relative expression analysis, knockout analysis, and cell treatment. These methods are employed to help ensure the reliability and specificity of our antibodies. At present, our portfolio comprises thousands of advanced verified antibodies. However, we remain committed to continuously expanding this range, as we strive to offer an even greater selection of high-quality antibodies to meet the diverse needs of our customers.

As shown in Figure 2, verification of GM130 Recombinant Rabbit Monoclonal Antibody (10H5L5) was performed using siRNA mediated knockdown of the target protein. With the help of immunofluorescence analysis, a significant reduction in GM130 expression was demonstrated in HeLa cells transfected with GM130 specific siRNA, confirming the specificity of the antibody.

Figure 2.
Knockdown of GM130 was achieved by transfecting HeLa cells with specific siRNA. Immunofluorescence analysis was performed on HeLa cells (untransfected, panel a-d), transfected with GM130 specific siRNA (panel i-l) or non-specific scrambled siRNA (panels e-h). Cells were fixed, permeabilized, and labeled with GM130 Recombinant Rabbit Monoclonal Antibody (10H5L5) (Cat. No. 703794, 1:100 dilution), followed by Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Cat. No. A32731, 1:2000 dilution). Nuclei (blue) were stained using ProLong Diamond Antifade Mountant with DAPI (Cat. No. P36962), and Rhodamine Phalloidin (Cat. No. R415, 1:300 dilution) was used for cytoskeletal F-actin (red) staining. Significant reduction of the signal was observed upon siRNA mediated knockdown (panel i-l) confirming the specificity of the antibody to GM130 (green).

Lot-to-lot consistency due to recombinant technology

Recombinant rabbit monoclonal antibodies are highly advantageous due to their excellent lot-to-lot reproducibility. This is primarily attributed to the fact that the production of these antibodies does not rely on traditional hybridoma cell lines, which are prone to a phenomenon known as cell drift. Cell drift refers to the gradual change in expression patterns exhibited by hybridoma cells over time, leading to variations in antibody production. In the case of recombinant rabbit monoclonal antibodies, the DNA sequences encoding the antibody's heavy and light chains are carefully banked and preserved. This means that the antibody production process does not involve the use of hybridoma cells, which are susceptible to changes in their expression patterns. As a result, recombinant rabbit monoclonal antibodies consistently maintain their initial specificity and performance characteristics across different production lots.

The example below demonstrates the specificity of SMAD2 Recombinant Rabbit Monoclonal Antibody (31H15L4) across four different lots through western blot. As seen in Figure 3, comparable antibody expression was observed in all lanes, indicating towards the consistency of the antibody. 

Improved antibody performance in multiple applications

Invitrogen antibodies undergo rigorous functional testing to help ensure their performance across a range of applications. Multiple well-established techniques, including western blot, immunocytochemistry, flow cytometry, and immunohistochemistry are employed to thoroughly assess the antibody's functionality and specificity. This comprehensive process helps ensure that our antibodies consistently deliver reliable and accurate results.

Figure 4 demonstrates testing of DYKDDDDK Tag Recombinant Rabbit Monoclonal Antibody (8H8L17), Alexa Fluor Plus 488 in multiple end-applications. This antibody has undergone thorough testing for western blot, flow cytometry, and immunocytochemistry applications. The successful validation across these multiple techniques enhances the confidence in the reliability and performance of this product.

Figure 4
. Demonstration of DYKDDDK tag antibody performance in multiple applications. A) Expression of DYKDDDDK Tag Recombinant Rabbit Monoclonal Antibody (8H8L17) (Cat. No. 701629RP488) shown in fluorescent western blot. Band of ~17kDa corresponding to DYKDDDDK tagged proteins was observed in HEK-293E transfected lysates on probing with the primary antibody. B) Flow cytometry analysis to confirm the expression of the same DYKDDDDK tag antibody. Pink histogram represents the expression profile of HEK-293E cells transiently transfected with HA-H3-DYKDDDDK as compared to untransfected cells (blue histogram). C) Immunofluorescent analysis of the DYKDDDDK tag antibody demonstrating the expression of the protein in panels a and d, compared to untransfected (panel e), DAPI (panel b), F-Actin (panel c), and no primary antibody controls (panel f).

Enhanced performance through engineering

Invitrogen recombinant rabbit monoclonal antibodies have reverse engineered IgG light and heavy chains. They are carefully developed to enhance the sensitivity of the antibodies, allowing for more accurate and reliable detection of target molecules. Furthermore, reverse engineered light and heavy chains also expand the application coverage of the antibodies, making them suitable for a wider range of research purposes. Importantly, while these enhancements are made, the specificity of the antibodies is not compromised. This means that researchers can confidently use our recombinant rabbit monoclonal antibodies knowing that they can achieve both high sensitivity and specificity in their experiments.

In Figure 5, performance of two recombinant rabbit monoclonal antibodies, Parkin Recombinant Rabbit Monoclonal Antibody (21H24L9) and OCT4 Recombinant Rabbit Monoclonal Antibody (3H8L6) has been compared to the wild type antibodies. The data clearly shows improved antibody performance upon engineering.

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* The use or any variation of the word “validation” refers only to research use antibodies that were subject to functional testing to confirm that the antibody can be used with the research techniques indicated. It does not ensure that the product(s) was validated for clinical or diagnostic use.

For Research Use Only. Not for use in diagnostic procedures.

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