SPCC417.12 Antibody

Introduction to SPCC417.12 Antibody

SPCC417.12 Antibody (product code CSB-PA527215XA01SXV) is a polyclonal antibody manufactured by Cusabio that specifically targets the SPCC417.12 protein from Schizosaccharomyces pombe strain 972 / ATCC 24843 (fission yeast) . As a research tool, this antibody enables the detection and analysis of the hypothetical esterase/lipase C417.12 protein, which has been identified through genomic sequencing but remains functionally uncharacterized .

The antibody is derived from rabbit immune systems and belongs to the IgG isotype class of immunoglobulins. It is designed for application in molecular and cellular biology research focusing on S. pombe, particularly for studies investigating esterase/lipase family proteins . The genomic location of the target gene has been identified at positions 1699739-1701301 in the S. pombe genome .

SPCC417.12 Protein Identity and Structure

The target of this antibody, SPCC417.12, is classified as an uncharacterized esterase/lipase (EC 3.1.-.-) belonging to the Type-B carboxylesterase/lipase family . This protein has been assigned the UniProt accession number O94493, indicating its registration in standardized protein databases . SPCC417.12 is thought to have a subcellular distribution primarily in the cytoplasm and nucleus of S. pombe cells .

Type-B carboxylesterases represent a distinct family of evolutionarily related proteins within the superfamily of proteins with the Alpha/beta hydrolase fold . These enzymes typically function through a catalytic triad consisting of three amino acid residues: serine, glutamate or aspartate, and histidine . This structure is evolutionarily conserved across various esterases and is crucial for their enzymatic activity.

pombe as a Model Organism

Understanding the context of S. pombe research is essential for appreciating the significance of the SPCC417.12 antibody. Schizosaccharomyces pombe has established itself as a valuable model organism in molecular and cellular biology research . Unlike its distant relative Saccharomyces cerevisiae (baker's yeast), which was initially studied to improve brewing and baking processes, S. pombe research has been driven by fundamental biological questions from its inception .

The foundations of S. pombe as a research model were established by Urs Leupold in the 1940s, who isolated the ancestor strains (968 h⁹⁰, 972 h⁻, and 975 h⁻) from which almost all modern laboratory strains derive . This genetic uniformity provides consistency in research data across different laboratories worldwide . The genome of S. pombe contains approximately 5,000 protein-coding genes, making it a relatively simple yet effective model for studying eukaryotic cellular processes .

S. pombe offers distinct advantages as a research model, including:

1. The ability to alternate between haploid and diploid states

2. A rod-shaped morphology that facilitates cell cycle studies

3. Conservation of many cellular processes found in higher eukaryotes

4. The presence of RNAi machinery components (Dicer, Argonaute, RNA-directed RNA polymerase)

These characteristics make S. pombe particularly valuable for studying gene regulation, cell division, and chromatin organization .

Antibody Production Process

SPCC417.12 Antibody is produced through immunization of rabbits with recombinant Schizosaccharomyces pombe SPCC417.12 protein . This process follows standard polyclonal antibody production protocols, where the immune system of the host animal (rabbit) generates a diverse array of antibodies against multiple epitopes on the target protein .

While the exact production method for this specific antibody is not detailed in the available data, the general process for recombinant antibody production at Cusabio involves several key steps as exemplified by their other antibody products:

1. Immunization of a host animal with a synthesized peptide or recombinant protein

2. Isolation of B cells from the immunized animal

3. RNA extraction and reverse transcription to cDNA

4. Extension of antibody genes using appropriate primers

5. Integration into expression vectors

6. Expression in host cells

7. Purification through affinity chromatography

The resulting SPCC417.12 antibody is purified using antigen affinity chromatography methods to ensure specificity and reduce non-target reactivity .

Physical and Chemical Properties

The SPCC417.12 Antibody is provided in liquid form with the following specifications:

Table 1: Physical and chemical properties of SPCC417.12 Antibody (CSB-PA527215XA01SXV)

The high glycerol content (50%) in the formulation serves as a cryoprotectant, preventing damage to the antibody during freeze-thaw cycles and extending shelf life . The preservative Proclin 300 (0.03%) inhibits microbial growth, while the PBS buffer at pH 7.4 maintains physiological conditions optimal for antibody stability .

Validated Applications

The SPCC417.12 Antibody has been validated for two primary applications:

1. ELISA (Enzyme-Linked Immunosorbent Assay): This application allows for quantitative detection of the target protein in solution .

2. Western Blot (WB): This technique enables detection of the target protein after separation by gel electrophoresis, providing information about molecular weight and relative abundance .

While these applications represent the validated uses of this antibody, the methodologies could potentially be extended to other experimental techniques commonly employed in antibody-based research, such as immunoprecipitation or immunohistochemistry, though these would require additional validation .

Western Blot Protocol for S. pombe Proteins

For Western blot applications using S. pombe proteins, researchers typically follow these general steps, which would apply to the SPCC417.12 Antibody:

1. Prepare S. pombe cell lysates (typically using glass bead disruption in appropriate lysis buffer)

2. Separate proteins by SDS-PAGE

3. Transfer proteins to a membrane (PVDF or nitrocellulose)

4. Block the membrane with appropriate blocking buffer

5. Incubate with SPCC417.12 Antibody (CSB-PA527215XA01SXV) at an optimized dilution

6. Wash the membrane

7. Incubate with an appropriate secondary antibody (typically anti-rabbit IgG)

8. Detect using a compatible visualization method

Pull-Down Experiments in S. pombe

The SPCC417.12 Antibody could potentially be used in pull-down/co-immunoprecipitation experiments to identify protein-protein interactions, following protocols similar to this:

1. Prepare S. pombe cell extracts in appropriate buffer

2. Add SPCC417.12 Antibody to the cell extract and incubate

3. Add protein A agarose beads to capture the antibody-protein complexes

4. Wash the beads to remove non-specific binding

5. Elute the bound proteins

6. Analyze by Western blot or mass spectrometry

This approach could help identify binding partners of the SPCC417.12 protein, potentially elucidating its function in S. pombe.

Manufacturing Quality Standards

The SPCC417.12 Antibody is subject to rigorous quality control measures that ensure its reliability in research applications. These quality controls include:

1. Purity Assessment: The antibody is guaranteed to exceed 90% purity as determined by SDS-PAGE analysis

2. ELISA Titer Determination: The antibody demonstrates a minimum ELISA titer of 1:64,000, indicating high sensitivity and specificity

3. Western Blot Validation: Positive detection of the antigen in Western blot applications is guaranteed

Value-Added Components

The SPCC417.12 Antibody product package includes additional components that enhance its research utility:

1. Positive Control: 200μg of antigen is provided, allowing for standardization and optimization of experimental conditions

2. Negative Control: 1ml of pre-immune serum is included, enabling proper experimental controls to be established

These additional components facilitate experimental design and troubleshooting, enhancing the reliability of research outcomes.

Complementary Research Tools

Researchers interested in studying the SPCC417.12 protein have access to several complementary tools that can enhance their investigations:

Table 2: Complementary research tools for SPCC417.12 protein studies

These recombinant proteins can serve as positive controls, standards for quantification, or as reagents for in vitro studies of protein function.

Database Resources for SPCC417.12 Research

Several database resources provide valuable information for researchers working with the SPCC417.12 protein:

1. KEGG database: The protein is registered under the identifier spo:SPCC417.12

2. UniProt database: The protein is registered under accession number O94493

3. Genome mapping resources: The gene is located at positions 1699739-1701301 in the S. pombe genome

These resources provide genomic context, protein sequence information, and potential functional annotations that can guide research design and interpretation.

Research Gaps and Challenges

Despite the availability of the SPCC417.12 Antibody as a research tool, several limitations exist in the current research landscape:

1. Limited Published Studies: There appears to be a scarcity of published research specifically utilizing the SPCC417.12 Antibody, limiting the availability of experimental protocols and optimization guidelines

2. Uncharacterized Target: The function of the SPCC417.12 protein remains largely uncharacterized, presenting challenges for experimental design and interpretation

3. Experimental Images: The manufacturer does not currently provide Western blot images, ELISA data, or immunofluorescence images for this specific antibody on their website, making it difficult for researchers to anticipate experimental outcomes

Future Research Opportunities

The SPCC417.12 Antibody presents several promising avenues for future research:

1. Functional Characterization: As a member of the Type-B carboxylesterase/lipase family, the SPCC417.12 protein likely plays a role in lipid metabolism or cellular signaling pathways. Studies using this antibody could help elucidate these functions

2. Protein-Protein Interaction Studies: Co-immunoprecipitation experiments using the SPCC417.12 Antibody could identify binding partners, potentially revealing functional networks

3. Subcellular Localization: Immunofluorescence studies could confirm and refine the predicted cytoplasmic and nuclear localization of the protein

4. Expression Analysis: Western blot analysis across different growth conditions or cell cycle stages could reveal regulatory patterns of SPCC417.12 expression