SLAIN1 Antibody is designed to bind specifically to the SLAIN1 protein (UniProt ID: Q8ND83), which belongs to the SLAIN motif-containing family. Key features include:
Parameter | Details |
---|---|
Immunogen | Recombinant human SLAIN1 protein (e.g., AA 43–136 or C-terminal peptide) |
Host | Rabbit (polyclonal) |
Reactivity | Human, mouse, and rat (varies by product) |
Form | Liquid (glycerol/PBS buffer with sodium azide) |
Purification | Affinity chromatography (antigen/protein G purified) |
Molecular Weight | Observed bands: 61–70 kDa (full-length), 34–40 kDa (isoforms/splice variants) |
The antibody is validated for multiple techniques:
Application | Recommended Dilution | Key Use Cases |
---|---|---|
Western Blot (WB) | 1:500–1:5,000 | Detection in Jurkat, HeLa, or A549 cell lysates |
ELISA | 1:2,000–1:10,000 | Quantitative analysis of SLAIN1 levels |
Immunohistochemistry (IHC) | 1:200–1:500 | Staining in paraffin-embedded brain or lung tissue |
Immunofluorescence (IF) | 1:20–1:200 | Localization in growth cones or cytoplasmic regions |
SLAIN1 regulates microtubule growth by recruiting polymerase ch-TOG to plus-ends, promoting persistent elongation . Disruption of SLAIN1-ch-TOG complexes increases catastrophe frequency and inhibits axon extension in neurons .
Idiopathic Pulmonary Fibrosis (IPF):
Neurodevelopment and Disease:
Cancer Research:
Antibody | WB Sensitivity | IF Specificity | Species Cross-Reactivity | Limitations |
---|---|---|---|---|
PACO57388 | High (61 kDa band) | Moderate | Human-only | Limited species coverage |
22123-1-AP | Moderate | High (EB1 colocalization) | Human/Mouse | Requires optimization in rodent models |
ABIN1450115 | Moderate | Moderate | Human/Mouse/Rat | Sodium azide preservative |
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.
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
These specifications are essential for selecting the appropriate antibody for specific experimental designs and determining compatibility with target samples.
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.
SLAIN1 expression has been confirmed in various cell types and tissues including:
This distribution suggests SLAIN1 may play roles in both specialized neural functions and general cellular processes common to diverse cell types.
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
Each application requires specific optimization parameters and has distinct advantages for different research questions.
The following dilution ranges are recommended for optimal results:
Application | Recommended Dilution |
---|---|
Western Blot (WB) | 1:500-1:1000 |
Immunofluorescence (IF)/ICC | 1:20-1:200 |
These are starting recommendations, and researchers should perform titration experiments to determine optimal concentrations for their specific experimental conditions .
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.
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.
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
Repeated freeze-thaw cycles should be avoided to maintain antibody performance and specificity.
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.
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.
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.
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.
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.
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.
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.