SS18L2 Antibody

What is SS18L2 antibody and what is its target protein?

SS18L2 antibody is a rabbit polyclonal antibody that specifically targets the SS18L2 (synovial sarcoma translocation gene on chromosome 18-like 2) protein. The antibody recognizes a specific immunogen sequence (VAFVPDWLRGKAEVNQETIQRLLEENDQLIRCIVEYQNKGRGNECVQYQHVLH) and is available as an affinity isolated antibody in buffered aqueous glycerol solution . SS18L2 is structurally related to SS18, which is involved in the pathognomonic translocation t(X;18)(p11;q11) found in synovial sarcoma, though SS18L2 itself has distinct functions and expression patterns .

What applications is SS18L2 antibody validated for?

SS18L2 antibody has been validated for multiple research applications including:

• Western Blot (WB): Recommended dilution 1:500-1:1000

• Immunohistochemistry (IHC): Recommended dilution 1:20-1:200

• ELISA

• Immunofluorescence: Recommended concentration 0.25-2 μg/mL

The antibody shows confirmed reactivity with human and mouse samples. For IHC applications, antigen retrieval with TE buffer pH 9.0 is suggested, with citrate buffer pH 6.0 as an alternative .

How does SS18L2 antibody differ from SS18-SSX fusion antibodies?

While SS18L2 antibody targets the SS18L2 protein, SS18-SSX antibodies specifically recognize the fusion protein resulting from the t(X;18) translocation in synovial sarcoma. SS18-SSX fusion-specific antibodies have been developed for diagnostic purposes with high sensitivity (95%) and specificity (100%) for synovial sarcoma . Unlike SS18-SSX fusion antibodies, which are primarily used for diagnostic purposes in cancer pathology, SS18L2 antibody is more commonly used in basic research investigating normal cellular processes and protein functions .

What are the optimal conditions for using SS18L2 antibody in immunohistochemistry?

For optimal IHC results with SS18L2 antibody:

• Section preparation: Use formalin-fixed, paraffin-embedded tissues sectioned at 4-6 μm thickness

• Antigen retrieval: Perform heat-mediated antigen retrieval using TE buffer pH 9.0 (optimal) or citrate buffer pH 6.0 (alternative)

• Dilution: Start with a 1:100 dilution and optimize based on your specific tissue sample

• Incubation: Overnight at 4°C or 60 minutes at room temperature

• Detection system: Use an appropriate detection system compatible with rabbit primary antibodies

• Controls: Include positive controls (human prostate cancer tissue has been validated) and negative controls

It's recommended to titrate the antibody in each testing system to obtain optimal results as sensitivity may be sample-dependent .

How should I optimize SS18L2 antibody for Western blot applications?

For optimal Western blot results:

Human liver tissue has been validated as a positive control for Western blot applications . Always include appropriate positive and negative controls, and consider running a gradient of antibody dilutions during optimization.

What methods can be used to validate the specificity of SS18L2 antibody?

To comprehensively validate SS18L2 antibody specificity:

• Knockout/knockdown validation: Compare results between wild-type samples and those with SS18L2 gene knockout or knockdown

• Peptide competition assay: Pre-incubate the antibody with excess immunizing peptide prior to application

• Multi-method validation: Confirm target detection using different methods (WB, IHC, IF)

• Cross-reactivity testing: Test against samples from different species to confirm species reactivity

• Mass spectrometry: Confirm the identity of immunoprecipitated proteins

• Antibody registry verification: Cross-check the antibody in validation databases

This approach is similar to how SS18-SSX fusion antibodies were validated using immunoblotting, immunoprecipitation, and chromatin immunoprecipitation followed by next-generation sequencing (ChIP-seq) .

How can SS18L2 antibody be utilized in chromatin immunoprecipitation (ChIP) experiments?

While not explicitly validated for ChIP in the search results, researchers can adapt SS18L2 antibody for ChIP experiments based on protocols established for similar antibodies like SS18-SSX:

• Crosslinking: Fix cells with 1% formaldehyde for 10 minutes at room temperature

• Chromatin preparation: Lyse cells and sonicate to generate 200-500 bp DNA fragments

• Pre-clearing: Pre-clear chromatin with protein A/G beads

• Immunoprecipitation: Incubate chromatin with SS18L2 antibody (3-5 μg) overnight at 4°C

• Washing: Perform stringent washes to remove non-specific binding

• Elution and reverse crosslinking: Elute protein-DNA complexes and reverse formaldehyde crosslinks

• DNA purification: Purify DNA for downstream applications

• Analysis: Analyze by qPCR, sequencing, or microarray

Similar antibodies have been successfully used to capture fusion proteins on chromatin at specific target sites and for genome-wide analysis .

What are the considerations for using SS18L2 antibody in co-immunoprecipitation studies?

For co-immunoprecipitation of SS18L2 and its interacting partners:

• Lysis buffer selection: Use a gentle lysis buffer (e.g., NP-40 buffer) to preserve protein-protein interactions

• Antibody amount: Use 2-5 μg of SS18L2 antibody per 500 μg of protein lysate

• Pre-clearing: Pre-clear lysates with protein A/G beads to reduce non-specific binding

• Binding conditions: Incubate antibody with lysate overnight at 4°C with gentle rotation

• Bead selection: Use protein A or A/G beads compatible with rabbit antibodies

• Washing stringency: Balance between maintaining interactions and reducing background

• Elution methods: Consider native elution with immunizing peptide or denaturing elution

• Controls: Include IgG control and input samples

As demonstrated with SS18-SSX antibodies, immunoprecipitation can be a powerful tool for studying protein interactions and complexes .

How does SS18L2 relate to the SS18-SSX fusion proteins in synovial sarcoma research?

While SS18L2 is related to SS18 (which forms the fusion with SSX in synovial sarcoma), they represent distinct proteins with different functions:

• Structural similarities: SS18L2 shares homology with SS18 but has unique domains and functions

• Expression patterns: Unlike the SS18-SSX fusion that is diagnostic for synovial sarcoma, SS18L2 is expressed in normal tissues

• Research applications: SS18-SSX antibodies are primarily used for diagnosis, while SS18L2 antibody is used to study normal cellular functions

• Cellular localization: Both proteins show nuclear localization, but may associate with different nuclear complexes

• Protein interactions: They likely have distinct interaction partners and contribute to different cellular processes

Understanding the relationship between these proteins can provide insights into both normal cellular functions and disease mechanisms .

What are common issues with SS18L2 antibody in IHC and how can they be resolved?

Common issues and solutions for SS18L2 antibody in IHC:

For human prostate cancer tissue, which has been validated as a positive control, proper antigen retrieval is particularly important for reliable results .

How should researchers interpret variable Western blot results with SS18L2 antibody?

When interpreting variable Western blot results:

• Band size verification: The expected molecular weight should be confirmed (verify against reported size for SS18L2)

• Multiple bands: May indicate post-translational modifications, splice variants, or proteolytic fragments

• Sample-specific variation: Expression levels may vary across tissue types and cell lines

• Loading controls: Always normalize to appropriate loading controls (β-actin, GAPDH)

• Species differences: Consider potential differences in protein size or epitope conservation between species

• Antibody lot variation: Document lot numbers and test new lots against previous ones

• Protocol consistency: Maintain consistent sample preparation and blotting protocols

Human liver tissue has been validated for Western blot applications with SS18L2 antibody and can serve as a reliable positive control .

What approaches can improve signal detection with SS18L2 antibody in low-expressing samples?

For enhancing detection in samples with low SS18L2 expression:

• Sample enrichment: Consider subcellular fractionation to concentrate the protein of interest

• Signal amplification: Implement tyramide signal amplification (TSA) for IHC/IF applications

• Increased exposure time: For Western blots, use longer exposure times with high-sensitivity ECL substrates

• Concentrated antibody: Decrease dilution within the recommended range (e.g., use 1:20 for IHC instead of 1:200)

• Enhanced antigen retrieval: Extend antigen retrieval time or try alternative buffers

• Sensitive detection methods: Use fluorescent secondary antibodies with signal accumulation capabilities

• Pre-treatment strategies: Treat cells with proteasome inhibitors to prevent protein degradation before analysis

• Alternative approaches: Consider RT-qPCR to confirm low expression at the mRNA level

These approaches should be systematically tested and optimized for your specific experimental system .

How does the performance of Prestige Antibodies like SS18L2 compare to standard antibodies?

Prestige Antibodies, including the SS18L2 antibody, offer several advantages over standard research antibodies:

• Extensive validation: Prestige Antibodies undergo thorough characterization through multiple methods and applications

• Documented specificity: They demonstrate low cross-reactivity due to stringent selection of antigen regions

• Affinity purification: This process enhances specificity by isolating only target-specific antibodies

• Comprehensive data access: All characterization data is accessible via the Human Protein Atlas portal

• Testing breadth: Each antibody is tested against tissue arrays of 44 normal human tissues and 20 common cancer types

• Cross-reactivity assessment: Protein arrays of 364 human recombinant protein fragments are used to evaluate specificity

• Reproducibility: Manufacturing processes emphasize lot-to-lot consistency

These features make Prestige Antibodies particularly valuable for research requiring high confidence in target specificity and reproducibility .

What advanced imaging techniques are most compatible with SS18L2 antibody immunofluorescence?

For advanced imaging with SS18L2 antibody:

• Confocal microscopy: Ideal for subcellular localization studies with optical sectioning capabilities

• Super-resolution microscopy:

  • STED (Stimulated Emission Depletion): For nanoscale visualization of protein distribution

  • STORM/PALM: For single-molecule localization at 20-30 nm resolution

• Multiphoton microscopy: For deep tissue imaging with reduced photobleaching

• Live-cell imaging: If used with membrane-permeable secondary antibody fragments

• FRET/FLIM: To study protein-protein interactions when combined with antibodies to interaction partners

• High-content screening: For automated analysis across multiple samples and conditions

• Correlative light and electron microscopy (CLEM): To correlate fluorescence with ultrastructural features

When using these advanced techniques, optimize fixation and antibody concentration (0.25-2 μg/mL is recommended) to maintain signal-to-noise ratio .

How is SS18L2 antibody used in cancer research compared to normal tissue studies?

SS18L2 antibody applications differ between cancer and normal tissue research:

In cancer research:

• Expression profiling: Evaluating SS18L2 expression across different tumor types

• Prognostic biomarker investigation: Correlating expression with clinical outcomes

• Tumor classification: Differentiating tumor subtypes based on expression patterns

• Treatment response monitoring: Assessing changes in expression following therapy

• Relationship to known oncogenic pathways: Studying interactions with established cancer-related proteins

In normal tissue studies:

• Tissue distribution mapping: Characterizing expression across normal human tissues

• Developmental biology: Studying expression changes during tissue development

• Physiological function investigation: Understanding normal biological roles

• Cell type specificity: Identifying cell populations expressing the protein

• Subcellular localization: Determining the protein's distribution within cellular compartments

The Human Protein Atlas project, which includes data from SS18L2 antibody testing, provides valuable information for both cancer and normal tissue research contexts .