egr2b Antibody

What is EGR2/KROX20 and why is it important for research?

EGR2 (Early Growth Response 2), also known as KROX20, is a zinc finger-containing transcription factor that belongs to the EGR C2H2-type zinc-finger protein family. The full-length protein has a molecular weight of approximately 50,302 daltons and exists in at least two identified isoforms . This protein plays crucial roles in multiple biological processes that make it a significant target for research investigations.

In neural development, EGR2 regulates hindbrain segmentation by controlling the expression of Hox genes, particularly HOXA4, HOXB2, and HOXB3, which helps specify odd and even rhombomeres during development . Within the peripheral nervous system, EGR2 is essential for myelination after birth, regulating the expression of myelin proteins like MPZ and promoting Schwann cell differentiation .

In the immune system, EGR2 plays important roles in T cell and NKT cell development and is predominantly expressed in the thymus . It is upregulated in T cells following TCR-crosslinking and may antagonize T cell activation while promoting apoptosis . Recent research has also identified EGR2 as a novel master transcriptional regulator of the INK/ARF locus and the senescence program .

At the molecular level, EGR2 functions as a sequence-specific DNA-binding transcription factor that binds to specific DNA sites in the promoter regions of various genes. It also acts as an E3 SUMO-protein ligase, facilitating SUMO1 conjugation to its coregulators NAB1 and NAB2, which in turn down-regulates EGR2's transcriptional activity .

How should I select between monoclonal and polyclonal EGR2 antibodies for my research?

Selecting between monoclonal and polyclonal EGR2 antibodies depends on your specific research needs and experimental applications. Both types offer distinct advantages that make them suitable for different research contexts.

Monoclonal EGR2 Antibodies

Monoclonal antibodies provide high specificity for a single epitope and consistent lot-to-lot reproducibility. They are excellent for detecting specific isoforms or post-translational modifications and ideal for applications requiring high precision like flow cytometry. For example, the erongr2 monoclonal antibody (available in PE and APC conjugated forms) is specific for Egr2 and does not cross-react with Egr3, making it ideal for distinguishing between related family members .

These antibodies are well-suited for experiments requiring consistent results over time, detection of specific EGR2 epitopes, and quantitative analyses. In flow cytometry applications, monoclonal antibodies like erongr2 have been tested by intracellular staining and flow cytometric analysis of stimulated mouse splenocytes using specialized buffer sets .

Polyclonal EGR2 Antibodies

Polyclonal antibodies recognize multiple epitopes on the EGR2 protein, potentially providing stronger signals, and are better for detecting denatured proteins. They are particularly useful for applications like Western blotting and immunohistochemistry. For instance, polyclonal antibodies like the rabbit polyclonal EGR2 antibody are generated against synthetic peptides corresponding to internal portions of the human EGR2 protein and can recognize multiple epitopes .

These antibodies are preferred for initial detection of EGR2 in new experimental systems, detection of EGR2 in fixed or denatured samples, and when signal amplification is necessary. The BSA-free polyclonal rabbit IgG anti-EGR2 antibody has been validated for multiple applications including ICC/IF, IHC, IHC-P, and Western blotting .

Selection Strategy

When choosing between these antibody types, consider:

• Your research question – whether you need to detect specific EGR2 domains/isoforms (choose monoclonal) or maximize detection sensitivity (choose polyclonal)

• Your application – Western blotting and IHC often work better with polyclonal antibodies; flow cytometry benefits from the specificity of monoclonals

• Species reactivity – verify the antibody's reactivity with your species of interest

• Validation data – review available data for your specific application and cell/tissue type

• Signal-to-noise ratio – consider the background staining typically observed with each antibody type

For cutting-edge research requiring both high specificity and sensitivity, consider using both antibody types in parallel experiments to provide complementary information and increase confidence in your results.

What are the optimal applications for different types of EGR2 antibodies?

EGR2 antibodies are versatile tools that can be used across a range of research applications, but certain antibody formulations are better suited for specific techniques. Understanding these optimal applications helps researchers select the most appropriate reagent for their experimental needs.

Western Blotting

For Western blotting, both monoclonal and polyclonal antibodies can be effective, but they require different optimization approaches:

• Polyclonal antibodies: Often provide stronger signals and can detect multiple epitopes even if some are lost during denaturation. For example, the rabbit polyclonal anti-EGR2 antibody (NB100-92327) has been validated for Western blot at dilutions of 1-2 μg/ml .

• Monoclonal antibodies: Provide high specificity but may be more sensitive to denaturation conditions. The recombinant monoclonal EGR2 antibody [EPR4004] has been validated for Western blotting at a 1/1000 dilution and successfully detects EGR2 in multiple cell lines including LnCaP, HepG2, MCF7, and SH-SY5Y .

When performing Western blots, expect to see a band at approximately 50 kDa, corresponding to the full-length EGR2 protein (50,302 daltons) .

Immunohistochemistry and Immunocytochemistry

For visualizing EGR2 in tissues and cells:

• Polyclonal antibodies: Such as NB100-92327 have been validated for IHC and ICC applications at concentrations of 2-5 μg/ml for IHC and 5-10 μg/ml for ICC .

• Application examples: In successful IHC applications, EGR2 antibodies typically reveal primarily nuclear staining in neurons and glial cells, although some cytoplasmic staining may also be observed, representing the translational or 14-3-3sigma interacting cytoplasmic pool of EGR2 .

Flow Cytometry

For flow cytometric analysis, fluorophore-conjugated monoclonal antibodies offer significant advantages:

• Conjugated antibodies: The erongr2 monoclonal antibody is available in both PE (12-6691-82) and APC (17-6691-82) conjugated forms, optimized for flow cytometry .

• Application protocols: These antibodies have been validated for intracellular staining followed by flow cytometric analysis, using specialized buffer sets like the Foxp3/Transcription Factor Staining Buffer Set .

• Dosage recommendation: These can be used at ≤0.25 μg per test, where a test is defined as the amount of antibody needed to stain a sample in a final volume of 100 μL .

Specialized Applications

• Immunoprecipitation: Some antibodies like the Early Growth Response Protein 2 Polyclonal Antibody (CAU25913) have been specifically validated for immunoprecipitation applications[9