Detect Calretinin in Tissue Samples?
Calretinin is a calcium-binding protein that belongs to the EF-hand family of proteins. It is encoded by the CALB2 gene. It is mainly found in the nervous system and in mesothelial cells (the cells lining body cavities such as the pleura, peritoneum, and pericardium).
The primary function of calretinin includes:
- Regulate calcium ion concentrations within cells.
- Help control neuronal excitability
- Signaling pathways
- Cell survival
In the nervous system, calretinin is found in subsets of GABAergic interneurons, where it modulates neuronal communication.
Outside the nervous system, calretinin expression is a diagnostic biomarker in pathology. For example, calretinin is strongly expressed in mesothelial cells. So, it acts as a valuable marker that helps distinguish malignant mesothelioma from other cancers such as adenocarcinoma.
Since it plays a critical role in the diagnosis of mesothelioma and other conditions, it is crucial to detect its amount accurately in tissue samples. Here is where CALB2 polyclonal antibodies come into play.
What are CALB2 Polyclonal Antibodies?
CALB2 polyclonal antibodies are laboratory-made antibodies that specifically recognize and bind to calretinin, a calcium-binding protein encoded by the CALB2 gene. These antibodies are produced by immunizing host animals (such as rabbits or goats) with calretinin protein or fragments of it.
The animal’s immune system generates a wide variety of antibodies that target different parts (epitopes) of the calretinin protein. The collected serum, which contains this mixture of antibodies, is then purified to create what we call CALB2 polyclonal antibodies.
These antibodies are known for their high sensitivity and versatility. So, they help detect even the smallest amount of calretinin in tissue samples.
How CALB2 Polyclonal Antibodies Detect Calretinin in Tissue Samples?
Calretinin is detected using various lab techniques. These techniques use antibodies that bind specifically to the protein of interest. Since CALB2 pAbs can bind to multiple epitopes of calretinin, they provide strong and reliable results across different techniques.
These techniques include:
Immunohistochemistry (IHC)
IHC (Immunohistochemistry) is used to detect calretinin in tissue samples. Calretinin is a protein found in neurons and mesothelial cells. Scientists and doctors can visualize the location and quantity of calretinin using IHC. It helps diagnose diseases like malignant mesothelioma.
In this technique:
- Tissue samples are preserved so cells and proteins stay intact.
- After this, the tissue is sliced into very thin layers to allow antibodies to reach calretinin.
- Now, a blocking solution is added to prevent antibodies from sticking to the wrong parts of the tissue.
- Now, the CALB2 polyclonal antibody is applied. It binds specifically to calretinin in the cells.
- Now, a enzyme-linked secondary antibody is added that binds to the primary antibody. As a result, it produces a visible signal. This signal shows where calretinin is in the tissue.
- Cells with calretinin appear colored under a microscope. Pathologists or researchers check which cells have calretinin. This helps in diagnosis and research.
Western Blotting (WB)
Like IHC, WB is another lab method used to detect calretinin protein in tissue samples. It is used to confirm the presence and size of calretinin. It is widely used to study neurons or mesothelioma cells.
In this method:
- The protein is extracted from tissue or cells.
- These proteins are placed on a gel. And then, an electric current is passed through the gel to separate them by size.
- After this, the proteins are moved from the gel onto a special membrane. This makes the detection process easier.
- Now, a blocking solution is added to prevent non-specific binding.
- CALB2 pAb is applied, which binds specifically to calretinin.
- Now, an enzyme-linked secondary antibody is added that produces a signal. This signal shows the position of calretinin on the membrane. This helps scientists confirm the presence and approximate size of the protein.
- The bands on the membrane indicate the amount of calretinin present in the sample, which researchers can compare to analyze the patient’s condition.
ELISA
ELISA stands for Enzyme-Linked Immunosorbent Assay. It is a plate-based technique used to detect the presence of calretinin in the tissue sample. It relies on antigen-antibody interactions.
In this technique:
- A special plate is coated with the sample containing calretinin.
- Now, CALB2 pAb is added, which binds to calretinin protein.
- Now, the plates are washed to remove excess antibodies. This helps prevent non-specific binding.
- Now, an enzyme-linked secondary antibody is added. After this, a substrate is added. The enzyme produces a signal by reacting with the substrate.
- The intensity of the signal is directly proportional to the amount of calretinin present in the sample. It is measured using a spectrophotometer.
The Bottom Line
CALB2 polyclonal antibodies are essential tools in both clinical diagnostics and scientific research. By binding to multiple epitopes of calretinin, they ensure sensitive, versatile, and reliable detection across different experimental techniques.
However, when using CALB2 pAbs for your experiments, make sure you buy them from a reliable source. Otherwise, you may have to compromise on your results!
