SMARCC1 Antibody

Structure and Function

SMARCC1 (SWI/SNF Related, Matrix Associated, Actin Dependent Regulator Of Chromatin Subfamily C Member 1), also known as BAF155, is a core subunit of the SWI/SNF chromatin remodeling complex . This 155 kDa protein is encoded by the SMARCC1 gene and contains a predicted leucine zipper motif typical of many transcription factors . SMARCC1 is intricately involved in transcriptional activation and repression of select genes by chromatin remodeling, specifically through alteration of DNA-nucleosome topology .

The protein functions as a component of SWI/SNF chromatin remodeling complexes that carry out key enzymatic activities, changing chromatin structure by altering DNA-histone contacts within a nucleosome in an ATP-dependent manner . This remodeling is essential for proper gene expression and cellular function, highlighting the importance of tools like SMARCC1 antibodies in studying these processes.

Role in Chromatin Remodeling Complexes

SMARCC1/BAF155 has been identified as a component of multiple SWI/SNF chromatin remodeling complexes including:

SMARCC1/BAF155 is one of the core subunits necessary for efficient nucleosome remodeling by BRG1 in vitro . Research suggests that it may stimulate the ATPase activity of the catalytic subunit of the complex , making it a crucial component for the functioning of these chromatin remodeling machines.

Developmental and Disease Implications

SMARCC1/BAF155 plays critical roles in development, particularly in neural development. During neural development, a switch from a stem/progenitor to a postmitotic chromatin remodeling mechanism occurs as neurons exit the cell cycle and become committed to their adult state . This transition requires a switch in subunit composition of the npBAF and nBAF complexes, with SMARCC1 being a constant component throughout this process.

The protein is an essential part of the mouse embryonic stem cell specific SWI/SNF complex (esBAF), which is necessary for early embryogenesis and proper brain and visceral endoderm development . Diseases associated with SMARCC1 include Hydrocephalus, Congenital, 5, and Congenital Hydrocephalus . Dysregulation of SMARCC1/BAF155 has also been linked to various diseases, including cancer, developmental disorders, and neurological conditions .

Polyclonal Antibodies

Polyclonal antibodies against SMARCC1/BAF155 are produced by immunizing animals (typically rabbits) with peptides or recombinant proteins corresponding to specific regions of the human SMARCC1 protein. These antibodies recognize multiple epitopes on the SMARCC1 protein, providing robust detection across various applications.

Several polyclonal SMARCC1 antibodies are commercially available:

• Active Motif's SMARCC1/BAF155 antibody (pAb), raised against a peptide within the N-terminal region of human SMARCC1/BAF155

• Abcam's Anti-SMARCC1/BAF155 antibody (ab264166), generated using a synthetic peptide within human SMARCC1 amino acids 300-350

• Abcam's Anti-SMARCC1/BAF155 antibody (ab126180), produced using a recombinant fragment protein within Human SMARCC1 amino acids 800-1100

• Cell Signaling Technology's SMARCC1/BAF155 Antibody #9053, which recognizes endogenous levels of total SMARCC1/BAF155 protein

These polyclonal antibodies offer broad epitope recognition, enhancing detection sensitivity in various experimental conditions.

Monoclonal Antibodies

Monoclonal antibodies against SMARCC1/BAF155 are derived from a single B-cell clone and recognize a specific epitope on the target protein. These antibodies offer high specificity and consistency between batches, making them valuable for applications requiring precise epitope recognition.

Notable examples include:

• HuaBio's SMARCC1/BAF155 Rabbit Monoclonal Antibody (Clone JB43-43), generated against a recombinant protein within human SMARCC1 amino acids 680-820

• Assay Genie's SMARCC1/BAF155 Rabbit Monoclonal Antibody (CAB4275), which targets a sequence corresponding to amino acids 700-800 of human SMARCC1/BAF155

Monoclonal antibodies provide excellent specificity for applications where distinguishing between closely related proteins or specific conformational states is critical.

Recombinant Antibodies

Recombinant antibodies represent the cutting edge of antibody technology, produced using recombinant DNA technology rather than traditional animal immunization methods. They offer advantages in terms of batch-to-batch consistency, ethical production, and the ability to engineer specific properties.

HuaBio's SMARCC1 Recombinant Rabbit Monoclonal Antibody [JB43-43] (ET7107-12) exemplifies this category, providing researchers with a highly reproducible research tool for studying SMARCC1 .

Immunogen Information

SMARCC1 antibodies are produced using various immunogens targeting different regions of the SMARCC1 protein:

The choice of immunogen affects the antibody's specificity, reactivity, and application suitability. Antibodies targeting conserved regions may demonstrate cross-reactivity with homologous proteins or across multiple species, while those targeting unique regions provide higher specificity.

Species Reactivity

SMARCC1 antibodies demonstrate varying reactivity with SMARCC1 proteins from different species:

This cross-reactivity information is crucial for researchers working with different model organisms, as it helps in selecting the appropriate antibody for specific experimental systems.

Applications in Research

SMARCC1 antibodies are versatile tools applicable in various research techniques:

These applications enable researchers to investigate SMARCC1 expression, localization, interactions, and functions in various biological contexts.

Recommended Protocols

For optimal results with SMARCC1 antibodies, manufacturers recommend specific protocols:

Western Blotting:

• The addition of 0.05% Tween 20 in the blocking buffer and primary antibody incubation buffer is recommended to aid in detection

• Using 5% Milk-TBST for blocking and as antibody diluent, with primary antibody incubation overnight

• For chromatin-bound proteins like SMARCC1 that may not be soluble in low salt nuclear extracts, a High Salt/Sonication Protocol is recommended for nuclear extract preparation

Immunofluorescence:

• Formaldehyde fixation of cells provides optimal results, as demonstrated with HeLa cells stained with SMARCC1/BAF155 antibody at a 1:2,000 dilution

Sample Preparation Considerations:

• Many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, a High Salt/Sonication Protocol is recommended when preparing nuclear extracts for Western blot analysis

• For immunoprecipitation followed by western blotting, using Light Chain specific secondary antibodies helps avoid heavy chain interference

Chromatin Remodeling Studies

SMARCC1 antibodies have been instrumental in elucidating the role of SMARCC1/BAF155 in chromatin remodeling processes. Research has shown that SMARCC1 is a core subunit of the SWI/SNF complex necessary for efficient nucleosome remodeling by BRG1 in vitro . These antibodies help researchers investigate how SMARCC1 contributes to the ATPase activity of the catalytic subunit of the complex .

The ability to detect and isolate SMARCC1-containing complexes has led to a better understanding of how these molecular machines regulate gene expression through chromatin structure alteration. By identifying the protein interactions and genomic binding sites of SMARCC1, researchers have gained insights into the mechanisms of transcriptional regulation mediated by SWI/SNF complexes.

Developmental Biology Research

SMARCC1 antibodies have been crucial in developmental biology studies, particularly in understanding neural development. Research has revealed that SMARCC1 is an essential part of the mouse embryonic stem cell specific SWI/SNF complex (esBAF), which is necessary for early embryogenesis and proper brain and visceral endoderm development .

Studies employing these antibodies have helped elucidate the role of SMARCC1 in the transition from neural progenitor cells to postmitotic neurons, which involves a switch in subunit composition of chromatin remodeling complexes . The npBAF complex, which contains SMARCC1, is essential for the self-renewal and proliferative capacity of multipotent neural stem cells, while the nBAF complex (also containing SMARCC1) plays a role in regulating genes essential for dendrite growth .

Disease-Related Research

The involvement of SMARCC1 in various diseases makes SMARCC1 antibodies valuable tools in pathological studies:

• Hydrocephalus: SMARCC1 has been linked to Hydrocephalus, Congenital, 5 and Congenital Hydrocephalus , and antibodies against SMARCC1 help study its role in this condition

• Cancer: Dysregulation of chromatin remodeling complexes, including SMARCC1-containing complexes, has been implicated in various cancers

• Neurological Disorders: Given SMARCC1's role in neural development, antibodies against this protein are valuable for studying neurological disorders associated with developmental abnormalities

• Developmental Disorders: Understanding the role of SMARCC1 in embryonic development provides insights into developmental disorders associated with chromatin remodeling defects

Selection Criteria for Research Applications

When selecting a SMARCC1 antibody for research, several factors should be considered:

1. Experimental Application: Different antibodies perform optimally in specific applications. For instance, some antibodies work well in western blotting but may not be suitable for immunohistochemistry or flow cytometry.

2. Species Reactivity: Ensure the antibody recognizes SMARCC1 from your species of interest. Cross-reactivity information is crucial for researchers working with different model organisms.

3. Epitope Location: Antibodies targeting different regions of SMARCC1 may give different results depending on protein conformation, post-translational modifications, or protein-protein interactions.

4. Validation Data: Review manufacturer-provided validation data, including western blots, immunofluorescence images, or other application-specific results to ensure the antibody performs as expected in your experimental setup.

5. Antibody Format: Consider whether the antibody format (polyclonal, monoclonal, or recombinant) is appropriate for your application. Polyclonal antibodies often provide higher sensitivity, while monoclonal antibodies offer greater specificity.

6. Buffer Compatibility: Check if the antibody buffer is compatible with your experimental conditions, particularly for specialized applications like live-cell imaging or enzymatic assays.

Emerging Applications

As research on chromatin remodeling continues to advance, new applications for SMARCC1 antibodies are emerging:

• Single-Cell Technologies: The integration of SMARCC1 antibodies into single-cell technologies to understand cell-to-cell variations in chromatin remodeling processes

• In Vivo Imaging: Development of fluorescently labeled SMARCC1 antibodies or antibody fragments for live-cell and in vivo imaging of chromatin dynamics

• Therapeutic Targets: Exploration of SMARCC1-targeting strategies as potential therapeutic approaches for conditions involving chromatin dysregulation

• Biomarker Development: Investigation of SMARCC1 as a potential biomarker for diseases associated with chromatin remodeling complex dysfunction

Technical Advancements

Ongoing advancements in antibody technology promise to enhance SMARCC1 antibody utility:

• Enhanced Specificity: Development of antibodies with higher specificity for particular SMARCC1 isoforms or post-translationally modified forms

• Improved Sensitivity: Creation of detection systems with higher sensitivity for low-abundance SMARCC1 complexes

• Multiparameter Analysis: Integration of SMARCC1 antibodies into multiplexed detection systems to simultaneously analyze multiple components of chromatin remodeling complexes

• Engineered Antibody Fragments: Development of smaller antibody fragments that can access restricted cellular compartments or recognize specific conformational states