stpg2 Antibody

What is STPG2 protein and why is it relevant for research?

STPG2, also known as Sperm-tail PG-rich repeat-containing protein 2 or C4orf37, is a human protein encoded by the STPG2 gene. The protein has a molecular weight of approximately 51 kDa as observed in Western blot analyses . While the specific function of STPG2 remains under investigation, it represents an important research target for understanding cellular processes related to its expression patterns. The protein is identifiable in the UniProt database under accession number Q8N412 .

The research interest in STPG2 stems from its potential involvement in cellular functions that may be relevant to both physiological and pathological processes. Antibodies against STPG2 enable researchers to investigate its expression, localization, and potential functional roles in various tissues and cell types.

What types of STPG2 antibodies are available for research applications?

Several types of STPG2 antibodies have been developed for research purposes:

• Polyclonal antibodies: Typically raised in rabbits against recombinant human STPG2 protein fragments. These recognize multiple epitopes of the STPG2 protein .

• Conjugated antibodies: Including FITC-conjugated polyclonal antibodies, which are particularly useful for immunofluorescence and flow cytometry applications .

Both types of antibodies are typically generated using recombinant human STPG2 protein fragments (such as amino acids 201-459) as immunogens .

How should researchers interpret Western blot results when using STPG2 antibodies?

When interpreting Western blot results with STPG2 antibodies, researchers should look for a band at approximately 51 kDa, which represents the predicted and observed size of the STPG2 protein . The appearance of multiple bands may indicate potential isoforms, degradation products, or non-specific binding.

To ensure accurate interpretation, researchers should:

• Always include positive controls (cells/tissues known to express STPG2) and negative controls (knockout cells or tissues)

• Use appropriate blocking agents to minimize background

• Optimize antibody concentration based on the manufacturer's recommendations (typically 1:1000-1:5000 dilution for Western blot)

• Consider using knockout validation approaches as demonstrated in systematic antibody validation studies

What are the optimal protocols for using STPG2 antibodies in Western blotting?

A standardized protocol for Western blotting with STPG2 antibodies should include:

Sample preparation:

• Prepare whole cell lysates (e.g., from HL60 cells as referenced in validation data)

• Include appropriate controls (positive samples, negative controls)

SDS-PAGE and transfer:

• Separate proteins using standard SDS-PAGE techniques

• Transfer to appropriate membrane (PVDF or nitrocellulose)

Antibody incubation:

• Block membrane with appropriate blocking solution

• Incubate with primary STPG2 antibody at 0.8μg/ml or diluted 1:1000-1:5000

• Wash thoroughly with TBST (Tris-buffered saline with 0.1% Tween-20)

• Incubate with appropriate secondary antibody (e.g., goat polyclonal to rabbit at 1/10000 dilution)

Detection and analysis:

• Use chemiluminescence or other detection methods

• Analyze band at expected 51 kDa molecular weight

This protocol aligns with validated methods shown to detect STPG2 in human cell lines .

How can STPG2 antibodies be effectively utilized in immunohistochemistry?

For optimal immunohistochemistry (IHC) results with STPG2 antibodies:

Tissue preparation:

• Use formalin-fixed, paraffin-embedded tissue sections

• Perform appropriate antigen retrieval (method may vary depending on fixation)

Staining procedure:

• Block endogenous peroxidase activity and non-specific binding

• Apply STPG2 antibody at a dilution of 1:20-1:200 (with 1:100 being validated for human spleen tissue)

• Incubate at appropriate temperature and duration (typically 4°C overnight)

• Apply suitable detection system based on primary antibody host species

Controls and validation:

• Include positive control tissues

• Include negative controls (primary antibody omission or isotype control)

• Consider using a systematic validation approach similar to those used for other antibodies

What systems biology approaches can enhance the analysis of data generated using STPG2 antibodies?

Systems biology approaches can provide comprehensive insights when working with antibody-generated data:

Network integration analysis:

• Integrate STPG2 expression data with public human blood transcriptomes

• Place findings in biological context using gene interaction data (interactome/bibliome)

Pathway enrichment analysis:

• Apply Gene Set Enrichment Analysis (GSEA) to identify molecular pathways associated with STPG2 expression

• Utilize databases such as Nature/NCI pathway interaction database for pathway level analyses

Modular analysis approaches:

• Consider using blood transcription modules (BTM) as alternatives to conventional pathway analyses

• Identify BTMs associated with STPG2 expression patterns

These approaches allow researchers to move beyond single-gene analysis to understand STPG2 in the context of broader biological systems and pathways .

How can researchers validate the specificity of STPG2 antibodies?

Validating antibody specificity is crucial for reliable research outcomes. For STPG2 antibodies, consider implementing:

Knockout validation approach:

• Compare antibody reactivity in wild-type vs. STPG2 knockout cell lines

• This approach has been successfully used for validating other antibodies like TGM2

Multi-application validation:

• Test antibody performance across multiple applications (WB, IHC, ELISA)

• Verify consistent results across applications using the same antibody

Epitope mapping:

• Determine the specific region of STPG2 recognized by the antibody

• Compare with known protein domains and potential cross-reactive epitopes

Peptide competition assays:

• Pre-incubate antibody with purified antigen peptide

• Verify signal reduction/elimination in subsequent experiments

A standardized experimental protocol comparing results in knockout cell lines against isogenic parental controls represents the gold standard for antibody validation .

What are common technical challenges when working with STPG2 antibodies and how can they be addressed?

Researchers commonly encounter several challenges when working with STPG2 antibodies:

Implementing these solutions can significantly improve experimental outcomes when working with STPG2 antibodies.

How can bioinformatics approaches be applied to analyze STPG2 antibody data in the context of broader immunological studies?

Bioinformatics provides powerful tools for analyzing STPG2 data within broader immunological contexts:

Principal Component Analysis (PCA) and Linear Discriminant Analysis (LDA):

• PCA can identify major patterns in STPG2 expression data across different tissues/conditions

• LDA can help classify samples based on STPG2 expression patterns when combined with other parameters

Integration with antibody polyreactivity databases:

• Compare STPG2 antibody characteristics with databases of antibody sequences

• Analyze physical and sequence-based properties to understand STPG2 antibody binding characteristics

Biophysical property analysis:

• Examine how hydrophobicity, charge, and structural parameters of the STPG2 antibody CDR loops influence specificity

• Apply computational tools to identify key determinants of antibody-antigen interactions

These approaches can reveal patterns that might not be apparent from traditional experimental analysis alone .

What are the optimal storage and handling conditions for STPG2 antibodies?

Proper storage and handling are essential for maintaining STPG2 antibody functionality:

Storage temperature:

• Upon receipt, store at -20°C or -80°C

• For FITC-conjugated antibodies, protect from light during storage

Aliquoting:

• Divide into small single-use aliquots to prevent repeated freeze-thaw cycles

• Clearly label each aliquot with antibody details and date

Buffer conditions:

• STPG2 antibodies are typically supplied in a storage buffer containing:

  • 0.03% Proclin 300 (preservative)

  • 50% Glycerol

  • 0.01M PBS, pH 7.4

Shipping and handling:

• Antibodies are typically shipped at 4°C

• Upon delivery, transfer immediately to recommended storage temperature

Adhering to these storage guidelines will maximize antibody shelf-life and experimental reproducibility.

How should researchers design appropriate controls for experiments using STPG2 antibodies?

Robust experimental controls are essential for reliable results with STPG2 antibodies:

Positive controls:

• Human cell lines known to express STPG2 (e.g., HL60 cells)

• Human tissues with documented STPG2 expression (e.g., spleen tissue)

Negative controls:

• Secondary antibody only (omit primary antibody)

• Isotype control (irrelevant primary antibody of same isotype)

• Ideally, STPG2 knockout cell lines or tissues (if available)

Technical controls:

• Loading controls for Western blot (housekeeping proteins)

• Tissue controls for IHC (internal positive and negative structures)

• Pre-absorption controls (antibody pre-incubated with immunizing peptide)

These controls help distinguish specific from non-specific signals and validate experimental findings, following the principles established in systematic antibody validation studies .

How can STPG2 antibodies be integrated into multi-parameter systems biology studies?

STPG2 antibodies can be valuable components of systems-level research:

Transcriptional signature analysis:

• Combine STPG2 protein expression data with transcriptomic analyses

• Identify coordinated expression patterns in specific biological contexts

Network integration:

• Place STPG2 in protein-protein interaction networks

• Identify potential functional relationships with other cellular components

Multi-omics integration:

• Correlate STPG2 protein levels (detected by antibodies) with transcriptomics, proteomics, and other omics data

• Apply computational frameworks for integrating heterogeneous datasets

This integration allows researchers to understand STPG2 function within broader biological systems rather than in isolation.

What methodological considerations are important when developing assays for detecting post-translational modifications of STPG2?

Detecting post-translational modifications (PTMs) of STPG2 requires careful methodological planning:

Antibody selection:

• Determine if standard STPG2 antibodies recognize modified forms

• Consider developing or sourcing modification-specific antibodies

Sample preparation:

• Include phosphatase inhibitors for phosphorylation studies

• Use appropriate lysis buffers that preserve the PTM of interest

Enrichment strategies:

• Consider immunoprecipitation with STPG2 antibodies followed by PTM-specific detection

• Use STPG2 antibodies validated for immunoprecipitation applications

Validation approaches:

• Implement controls similar to those used in antibody validation studies

• Include treatments that modulate the PTM of interest as positive/negative controls

These methodological considerations will enhance the reliability of assays designed to detect STPG2 modifications.