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17 January, 2024 by Anshul (neobio)
Are you tired of being overwhelmed by the quantities of options for antibodies for Influorescence, Western Blotting, Flow Cytometry, or Immunohistochemistry (IHC)? Well, you’re not alone. As a research scientist in a top institution, finding the ideal, reliable, and highly specific monoclonal antibodies for your research can feel daunting. That’s why we here at NeoBiotechnologies have determined to take you through a detailed review of the 5 best antibodies for IHC.
IHC is a powerful technique used to detect and localize specific antigens in cells and tissue, echoing the unique bond between an antibody and an antigen. This tool performs optimally for research settings, and its value in the clinical diagnostics landscape is continually increasing. An appreciation of the IHC process, compatible with formalin-fixed, paraffin-embedded (FFPE) tissue and automated methods for reproducibility, can aid in the astute selection of antibodies for IHC.
In the mainframe of IHC, the participation of antibodies is crucial. These are typically targeted against certain antigens present in specific tissues and cells, which assists in determining the cellular type and origin of the organ. In the grand scheme of immunological reactions, these antibodies circle the bloodstream or are attached to the surface of lymphocytes. Here, they specialize in interacting specifically with foreign antigens, leading to their ingestion.
In IHC, there are two primary detection protocols – direct and indirect. The direct detection method employs labeled primary antibodies to target antigens. On the other hand, indirect detection necessitates using secondary antibodies, key for signal amplification, since more than one secondary antibody molecule can bind to each primary antibody.
Both primary and secondary antibodies play integral roles in IHC. Primary antibodies acknowledge and bind directly to the target antigen, while secondary antibodies recognize primary antibodies. The appropriate selection and usage of these antibodies incredibly influence the success of your IHC experiments.
In conclusion, here are some key points about the antibodies for IHC:
To wrap things up, here’s an infographic that provides some quick answers for anyone searching for ‘antibodies for IHC’.
When it comes to selecting the right antibodies for Immunohistochemistry (IHC), there are several critical factors to take into account. Let’s delve into each one of them.
The first step in choosing an antibody for IHC is defining your protein target of interest. Protein targets can be complex, given their diverse nature and structures. It’s crucial to understand your target, its function, and its location within a cell. This information will guide your selection of the most appropriate antibody that can bind specifically and effectively to your target.
The next factor to consider is the compatibility of the antibody with your sample. Antibodies vary in their sequence and structure, which can affect their ability to recognize and bind to target proteins. This is particularly important when working with samples from different species, as the same protein can have slight variations in its sequence across species. Therefore, always ensure that the chosen antibody is compatible with the species and tissue type of your sample.
The choice between monoclonal and polyclonal antibodies is another crucial decision. Monoclonal antibodies bind to a single epitope, offering high specificity. Polyclonal antibodies, on the other hand, recognize multiple epitopes on a single target protein, offering greater sensitivity and often more stability over a range of pH and salt concentrations. For these reasons, polyclonal antibodies are often preferred for IHC.
Although both types have their benefits, at NeoBiotechnologies, we manufacture over 1,000 highly validated, monospecific Rabbit Recombinant Monoclonal Antibodies, ideal for IHC, Flow Cytometry, Western Blotting, or Immunofluorescence.
Lastly, the process of antigen retrieval (AR) is vital in IHC. AR involves the pretreatment of tissue to retrieve antigens masked by fixation, making them more accessible to antibody binding. This step significantly increases the sensitivity of IHC and expands its application. The method of AR depends on the specific target antigen and antibody. Thus, choosing an antibody that is compatible with the AR method you intend to use can greatly influence the success of your IHC experiment.
In conclusion, choosing the right antibodies for IHC involves careful consideration of the target protein, compatibility of the antibody with the sample, type of antibody, and the role of antigen retrieval. Making an informed decision in these areas will set the foundation for successful IHC experiments.
In this section, we delve into the specifics of the top 5 antibodies for IHC, highlighting their features, benefits, and applications. At NeoBiotechnologies, we manufacture over 1,000 highly validated, monospecific Rabbit Recombinant Monoclonal Antibodies, perfect for Immunohistochemistry, Flow Cytometry, Western Blotting, or Immunofluorescence.
This antibody is a powerful tool for detecting claudin 5 in kidney tissue. One of its key features is its ability to show staining in the membrane and cytoplasm of paraffin-embedded human kidney tissue. It’s used at a dilution of 1:20 and has been successfully involved in antigen retrieval performed using 10mM sodium citrate.
The Cardiac Troponin T Mouse Monoclonal Antibody excels in analyzing mouse heart tissue. It’s used at a concentration of 5µg/mL, with an overnight incubation at 4°C. This antibody allows for the detection of Cardiac Troponin T (green) in heart longitudinal sections.
Cyclin E is part of the highly conserved cyclin family, and this antibody is ideal for studying its role in the regulation of CDK kinases. It is particularly useful for observing the expression and degradation patterns of Cyclin E, which contributes to the temporal coordination of each mitotic event.
CD34 is a well-known marker for hematopoietic stem cells, and our CD34 Mouse Monoclonal Antibody is a powerful tool for its detection. Its high specificity and sensitivity make it an excellent choice for IHC studies of these crucial cells.
Our Keratin 7 Rabbit Polyclonal Antibody is a versatile tool for studying the keratin family of intermediate filament proteins. It’s particularly useful in IHC studies for detecting epithelial cells, as keratin 7 is often expressed in these cells.
In conclusion, the optimal antibodies for IHC will depend on the specific needs of your research. Each of these antibodies offers unique benefits and applications, making them excellent choices for various IHC studies. At NeoBiotechnologies, we are committed to providing highly validated and specific monoclonal antibodies to aid your research.
The success of Immunohistochemistry (IHC) experiments heavily depends on the optimization of antibodies. This involves adjusting various factors such as incubation time and temperature, dealing with specific staining and background signal, and handling high-affinity antibodies’ concentration and incubation. In this section, we will delve into these aspects to provide you with a better understanding of how to optimize antibodies for IHC.
The incubation time and temperature play a significant role in the success of IHC. They influence the binding of the primary antibody to the target antigen. It’s important to find the optimal balance, as too short an incubation time may lead to insufficient binding, while too long can cause over-staining or high background signals.
Usually, a longer incubation time at a lower temperature (for instance, overnight at 4°C) is preferred over a shorter period at room temperature. However, these conditions can vary based on the specific antibody and antigen involved in your study. Thus, it’s recommended to titrate different antibody dilutions while keeping the incubation time and temperature constant to achieve the best results.
Achieving specific staining with minimal background signal is crucial for clear and accurate IHC results. If you’re obtaining specific staining but also experiencing a high background signal, you might need to adjust the incubation time and temperature again.
Additionally, consider the blocking step. This process prevents non-specific binding of the antibodies to proteins other than the target antigen, which can contribute to the background signal. The choice of the blocking agent can also influence the background, and it’s worth experimenting with different agents to see which one gives the lowest background for your specific situation.
High-affinity antibodies, such as the Rabbit Recombinant Monoclonal Antibodies we manufacture at NeoBiotechnologies, bind to their target antigens with a very high affinity. This means they can be applied at high dilutions, thereby reducing the chance of non-specific interactions.
However, very high dilution may also lead to signal attenuation, especially where incubation times are limited. In such cases, antibody amplification systems can be employed to overcome this challenge. Though, that while these systems allow a higher dilution of the primary antibody (and hence higher specificity), they could also amplify non-specific signals if not properly controlled.
In conclusion, the optimization of antibodies for IHC is a careful balancing act that requires attention to several key factors. But with patience and careful experimentation, you can achieve clear, specific staining that will provide valuable insights into your protein target of interest. At NeoBiotechnologies, we are committed to providing researchers like you with the tools and support you need to make this process as straightforward as possible.
We’ve walked through the top 5 antibodies for IHC, each of them offering unique features and benefits. These antibodies have been carefully selected for their high specificity, reliability, and versatility across various applications. From defining your protein target to ensuring compatibility with your sample, these antibodies cater to a wide range of needs in the IHC field.
Selecting the right antibodies for IHC isn’t a one-size-fits-all process. It involves a keen understanding of your protein target, rigorous testing, and thoughtful consideration of both the antibody and your sample. With the right antibody, IHC can become a powerful tool in your research, enabling you to visualize and understand the distribution and localization of specific proteins within a cell or tissue sample.
Polyclonal antibodies offer stability across a range of pH and salt concentration, making them frequently used in IHC experiments. However, monoclonal antibodies provide specificity that can be advantageous in certain applications. The choice between these two types will depend on your specific needs and the nature of your research.
At NeoBiotechnologies, we understand the importance of reliable and highly validated antibodies for IHC. We’ve taken the time to manufacture and validate over 1,000 monospecific Rabbit Recombinant Monoclonal Antibodies, ideal for various applications such as Immunohistochemistry, Flow Cytometry, Western Blotting, or Immunofluorescence.
Our commitment to quality and precision ensures that the antibodies we offer are not just products – they’re tools for advancement in the scientific community. We strive to provide antibodies that can be trusted, allowing researchers like you to focus on the critical task of making discoveries that push the boundaries of our understanding of biology and disease.
With NeoBiotechnologies, you’re not just choosing antibodies. You’re choosing a partner dedicated to supporting your research and helping you achieve your goals. We’re here to provide the antibodies for IHC you need to move your research forward.