SLAIN1 Antibody

What is SLAIN1 and what is its biological significance?

SLAIN1 (SLAIN motif family, member 1) is a protein encoded by the SLAIN1 gene (Gene ID: 122060). It belongs to the SLAIN motif family and has a calculated molecular weight of approximately 61 kDa, though it is often observed at 65-70 kDa and 34-40 kDa in experimental conditions . While the complete function of SLAIN1 remains under investigation, it contributes to cellular processes that make it a valuable target for various research applications. The protein's specific motif structures and interactions with other cellular components make it relevant for studies in normal cellular physiology and potential disease states.

What are the critical technical specifications of SLAIN1 antibodies?

SLAIN1 antibodies are typically generated as polyclonal antibodies in rabbits using SLAIN1 fusion proteins as immunogens. Key specifications include:

• Host/Isotype: Rabbit/IgG

• Reactivity: Proven reactivity with human and mouse samples

• Applications: Western Blot (WB), Immunofluorescence (IF), Immunocytochemistry (ICC), and ELISA

• Form: Liquid in PBS buffer with 0.02% sodium azide and 50% glycerol at pH 7.3

• Purification method: Antigen affinity purification

These specifications are essential for selecting the appropriate antibody for specific experimental designs and determining compatibility with target samples.

Why does SLAIN1 show different molecular weights in experimental conditions?

SLAIN1 has a calculated molecular weight of 61 kDa but is typically observed at 65-70 kDa and sometimes at 34-40 kDa in laboratory conditions . This discrepancy may result from:

• Post-translational modifications such as glycosylation, phosphorylation, or other covalent additions

• Alternative splicing generating different isoforms

• Proteolytic processing creating truncated versions of the protein

• Experimental conditions affecting protein migration in gel electrophoresis

Researchers should anticipate these multiple bands when performing Western blot analysis and validate band identity through appropriate controls.

In which cell types has SLAIN1 expression been confirmed?

SLAIN1 expression has been confirmed in various cell types and tissues including:

• HeLa cells

• HEK-293 cells

• Mouse brain tissue

This distribution suggests SLAIN1 may play roles in both specialized neural functions and general cellular processes common to diverse cell types.

What are the validated applications for SLAIN1 antibodies?

SLAIN1 antibodies have been validated for the following applications:

• Western Blot (WB): For protein quantification and molecular weight determination

• Immunofluorescence (IF)/Immunocytochemistry (ICC): For subcellular localization studies

• ELISA: For quantitative detection in solution

Each application requires specific optimization parameters and has distinct advantages for different research questions.

What dilution ranges are recommended for different applications?

The following dilution ranges are recommended for optimal results:

These are starting recommendations, and researchers should perform titration experiments to determine optimal concentrations for their specific experimental conditions .

What is the optimal protocol for Western blot detection of SLAIN1?

The Western blot protocol for SLAIN1 detection should follow these key steps:

• Sample preparation: Lyse cells or tissues in appropriate buffer with protease inhibitors

• Protein separation: Use 10-12% SDS-PAGE gels for optimal resolution

• Transfer: Transfer proteins to PVDF or nitrocellulose membranes

• Blocking: Block with 5% non-fat milk or BSA in TBST buffer

• Primary antibody: Apply SLAIN1 antibody at 1:500-1:1000 dilution overnight at 4°C

• Washing: Wash membranes 3-5 times with TBST

• Secondary antibody: Apply appropriate HRP-conjugated secondary antibody

• Development: Visualize using ECL detection system

Expected results include bands at approximately 65-70 kDa and possibly at 34-40 kDa . Each laboratory should optimize these parameters based on their specific equipment and samples.

What considerations are important for immunofluorescence detection of SLAIN1?

For optimal immunofluorescence (IF) detection of SLAIN1:

• Fixation method: Use 4% paraformaldehyde for 15-20 minutes at room temperature

• Permeabilization: Permeabilize with 0.2-0.5% Triton X-100 for 5-10 minutes

• Blocking: Block with 1-5% BSA or normal serum for 30-60 minutes

• Primary antibody: Apply SLAIN1 antibody at 1:20-1:200 dilution for 1-2 hours at room temperature or overnight at 4°C

• Washing: Wash cells 3-5 times with PBS

• Secondary antibody: Apply fluorophore-conjugated secondary antibody

• Counterstaining: Use DAPI for nuclear staining

• Mounting: Mount slides with anti-fade mounting medium

SLAIN1 antibodies have been successfully used to detect the protein in HeLa and HEK-293 cells , providing valuable information about subcellular localization and expression patterns.

What are the optimal storage conditions for SLAIN1 antibodies?

SLAIN1 antibodies should be stored at -20°C for long-term stability. The antibodies are reported to be stable for one year after shipment when stored properly. For the specific product referenced in the search results:

• Aliquoting is unnecessary for -20°C storage

• The storage buffer contains PBS with 0.02% sodium azide and 50% glycerol at pH 7.3

• Some preparations may contain 0.1% BSA as a stabilizer

Repeated freeze-thaw cycles should be avoided to maintain antibody performance and specificity.

What are common causes of false negative results when using SLAIN1 antibodies?

False negative results can occur due to several factors:

• Insufficient protein in samples (verify total protein concentration)

• Protein degradation (ensure proper sample handling and add protease inhibitors)

• Inefficient protein extraction (optimize lysis buffer composition)

• Improper antibody dilution (titrate to determine optimal concentration)

• Suboptimal incubation conditions (verify temperature and duration)

• Issues with detection systems (include positive controls)

• Expression levels below detection threshold (consider enrichment methods)

Systematic troubleshooting of each step in the experimental workflow can help identify and address the source of false negative results.

How can researchers validate the specificity of SLAIN1 antibodies?

Validation of SLAIN1 antibody specificity should employ multiple approaches:

• Positive control samples: Use cells known to express SLAIN1 (HeLa, HEK-293 cells, or mouse brain tissue)

• Blocking peptide competition: Pre-incubate antibody with immunizing peptide to confirm specific binding

• Knockdown/knockout validation: Compare signals in SLAIN1-depleted vs. normal samples

• Multiple antibodies: Use antibodies targeting different epitopes of SLAIN1

• Predicted molecular weight verification: Confirm band pattern at expected molecular weights (61-70 kDa and potentially 34-40 kDa)

• Orthogonal techniques: Correlate antibody results with mRNA expression data

These approaches collectively provide strong evidence for antibody specificity and reliability.

What controls should be included when working with SLAIN1 antibodies?

Essential controls for SLAIN1 antibody experiments include:

• Positive tissue/cell controls: Samples known to express SLAIN1 (e.g., HeLa cells, HEK-293 cells, mouse brain tissue)

• Negative controls: Samples with minimal SLAIN1 expression or SLAIN1-knockout samples

• Loading controls: Housekeeping proteins (β-actin, GAPDH, tubulin) to normalize expression levels

• Secondary antibody-only control: To detect non-specific binding of secondary antibody

• Isotype control: Non-relevant rabbit IgG to identify non-specific binding

• Peptide competition control: Pre-incubation with immunizing peptide to verify specificity

These controls help distinguish specific from non-specific signals and validate experimental findings.

What is known about SLAIN1's role in disease states?

While comprehensive disease associations for SLAIN1 are still emerging, there is evidence of its potential relevance in certain pathological conditions. One study identified SLAIN1 as a potential biomarker in idiopathic pulmonary fibrosis through machine learning analysis . This suggests that altered SLAIN1 expression or function may contribute to disease processes involving tissue remodeling or repair. Further research is needed to fully characterize SLAIN1's role in both normal physiology and pathological states across different tissue types and disease models.

How can researchers effectively study SLAIN1 interactions with other proteins?

To investigate SLAIN1 protein interactions, researchers can employ several complementary approaches:

• Co-immunoprecipitation (Co-IP): Using SLAIN1 antibodies to pull down protein complexes

• Proximity ligation assay (PLA): For visualizing protein interactions in situ

• Yeast two-hybrid screening: To identify novel interaction partners

• Mass spectrometry following immunoprecipitation: For unbiased identification of interaction partners

• FRET/BRET analysis: For studying dynamics of protein interactions in living cells

• GST pull-down assays: Using recombinant SLAIN1 to identify direct binding partners

When designing these experiments, researchers should consider the potential impact of detergents and buffer conditions on preserving protein-protein interactions.

What approaches can address cross-reactivity concerns with SLAIN1 antibodies?

Cross-reactivity concerns can be addressed through:

• Epitope sequence analysis: Compare the immunizing sequence against proteome databases to identify potential cross-reactive proteins

• Validation in knockout/knockdown systems: Test antibody performance in SLAIN1-depleted samples

• Peptide competition assays: Determine if specific peptides can block antibody binding

• Testing in multiple species: Compare reactivity patterns across species with known sequence differences

• Multiple antibody validation: Use antibodies against different SLAIN1 epitopes to confirm findings

• Orthogonal methods: Correlate protein detection with RNA expression data

For SLAIN1 specifically, antibodies have shown reactivity with both human and mouse samples , suggesting conservation of epitopes across these species.

How can SLAIN1 antibodies be integrated into multi-omics research approaches?

Integration of SLAIN1 antibody-based detection with other omics approaches can provide comprehensive insights:

• Proteogenomics: Correlate SLAIN1 protein levels with genomic and transcriptomic data

• Phosphoproteomics: Identify post-translational modifications affecting SLAIN1 function

• Interactomics: Map the SLAIN1 protein interaction network using antibody-based pulldowns

• Spatial proteomics: Determine subcellular localization using immunofluorescence in conjunction with organelle markers

• Single-cell analysis: Combine antibody-based detection with single-cell RNA sequencing data

• Functional genomics: Correlate SLAIN1 protein levels with phenotypic changes following genetic manipulation

This multi-dimensional approach can reveal functional relationships and regulatory mechanisms governing SLAIN1 activity in normal and pathological contexts.