HSPA1L Monoclonal Antibody

What is HSPA1L and what cellular functions does it perform?

HSPA1L belongs to the heat shock protein 70 (HSP70) family, functioning as a molecular chaperone with a molecular weight of approximately 70 kDa. It plays crucial roles in protein folding, stress response, and cellular protection mechanisms. According to recent findings, HSPA1L forms complexes with other proteins like vaspin and contributes to protecting proximal tubular cells from stress-induced damage. This protein-protein interaction appears to ameliorate endoplasmic reticulum stress, autophagy impairment, and lysosome dysfunction in various disease models . Understanding HSPA1L's function is essential for designing experiments targeting stress response pathways.

What applications are suitable for HSPA1L monoclonal antibodies?

HSPA1L monoclonal antibodies can be utilized across multiple applications with specific dilution recommendations:

• Western Blotting (WB): 0.01-3 μg/mL

• Immunohistochemistry (IHC): 5-30 μg/mL

• Immunocytochemistry/Immunofluorescence (ICC/IF): 5-30 μg/mL

• Immunoprecipitation (IP): 0.5-4.0 μg for 1.0-3.0 mg of total protein lysate (based on polyclonal antibody protocols)

• ELISA applications

Researchers should note that optimal working dilutions must be determined by the end-user for specific experimental systems to achieve optimal signal-to-noise ratios .

What species reactivity is confirmed for HSPA1L monoclonal antibodies?

Current HSPA1L monoclonal antibodies have confirmed reactivity with human samples as specified in product documentation . For polyclonal antibodies against HSPA1L, reactivity has been demonstrated with human, mouse, and rat samples . When planning cross-species studies, researchers should verify the specific reactivity profile of their selected antibody, as cited reactivity may include human, mouse, rat, and bovine samples depending on the antibody preparation .

What is the role of HSPA1L in disease models and pathology?

HSPA1L has emerging significance in disease pathology, particularly in kidney disease models. Research indicates that HSPA1L forms a complex with vaspin that protects proximal tubular cells through multiple mechanisms:

• Amelioration of endoplasmic reticulum stress

• Reduction of autophagy impairment

• Prevention of lysosome dysfunction in diabetic kidney disease

Furthermore, genetic studies have identified rare, likely damaging HSPA1L variants associated with spontaneous preterm birth (SPTB) through whole exome sequencing, suggesting its role in reproductive outcomes . When designing disease model studies, consider that HSPA1L functioning may be tissue and condition-specific.

How does the vaspin-HSPA1L complex mechanism function at the cellular level?

The vaspin-HSPA1L complex demonstrates a protective mechanism in proximal tubular cells through clathrin-mediated endocytosis. This internalization process ameliorates obesity-induced lysosomal membrane permeabilization, inflammasome activation, and autophagy failure . Research has shown that albumin overload in proximal tubular cells increases HSPA1L secretion into the culture medium, resulting in reduced intracellular HSPA1L levels. This mechanism suggests how albuminuria disrupts HSPA1L-dependent organellar function in diabetic kidney disease . When investigating this pathway, co-immunoprecipitation assays can confirm protein interactions between vaspin and HSPA1L.

What genetic variants of HSPA1L have functional significance in human disease?

Several rare HSPA1L variants have been identified with potential functional impact:

• rs34620296 and rs150472288: Identified in a discovery cohort

• rs482145 and rs139193421: Identified in a replication cohort

These variants have been predicted as damaging by multiple in silico tools including SIFT, PolyPhen-2, MutationTaster, and MutationAssessor . According to Combined Annotation Dependent Depletion (CADD) scores, three of these variants (excluding rs139193421) rank among the top 1% of deleterious variants in the human genome . When conducting genetic association studies involving HSPA1L, researchers should consider these known functional variants.

What are the optimal storage conditions for HSPA1L monoclonal antibodies?

HSPA1L monoclonal antibodies require specific storage conditions to maintain stability and functionality:

• For frequent use: Store at 4°C

• For long-term storage: Aliquot and store at -20°C for up to 24 months

• Avoid repeated freeze/thaw cycles that can degrade antibody quality

Stability testing through accelerated thermal degradation (37°C for 48h) has demonstrated less than 5% loss rate within the expiration date under appropriate storage conditions . Researchers should monitor antibody performance over time, particularly when using older antibody preparations.

What controls should be included when using HSPA1L antibodies?

When designing experiments with HSPA1L antibodies, include appropriate controls:

These controls are essential for publication-quality data and ensuring the validity of experimental findings regarding HSPA1L expression or interactions.

What antigen retrieval methods are recommended for HSPA1L immunohistochemistry?

For optimal HSPA1L detection in immunohistochemistry applications:

• Primary recommendation: Antigen retrieval with TE buffer pH 9.0

• Alternative method: Antigen retrieval with citrate buffer pH 6.0

These retrieval methods have been validated for detection in various tissues including human endometrial cancer tissue, human breast cancer tissue, and mouse brain tissue . Researchers should optimize retrieval conditions based on specific tissue characteristics and fixation methods to maximize signal intensity while minimizing background.

How can researchers troubleshoot weak or non-specific signals with HSPA1L antibodies?

When encountering signal issues with HSPA1L antibodies, consider these troubleshooting approaches:

Remember that antibody titration is recommended in each testing system to obtain optimal results, as performance can be sample-dependent .

What methods can be used to validate HSPA1L-protein interactions?

To validate HSPA1L interactions with other proteins like vaspin or GRP78:

• Co-immunoprecipitation (Co-IP): Demonstrated effective for validating HSPA1L interactions in HEK293T cells with overexpression systems

• Immunofluorescence co-localization: Effective using HSPA1L antibodies (20μg/ml) with FITC-linked secondary antibodies (1μg/ml)

• Proximity ligation assays: Can provide in situ validation of protein interactions

• Western blot analysis following IP: Confirms specific protein interactions using antibody dilutions of 1:500-1:2000

When reporting protein interactions, include appropriate controls and quantitative measures of co-localization or interaction strength for comprehensive analysis.

What is the validated detection profile of HSPA1L monoclonal antibodies?

HSPA1L monoclonal antibodies have been validated in multiple sample types with specific detection characteristics:

This validation profile enables researchers to select appropriate positive controls and anticipate expected results in their experimental systems.

How do different HSPA1L antibody preparations compare in sensitivity and specificity?

When comparing monoclonal versus polyclonal HSPA1L antibody preparations:

Monoclonal antibodies (e.g., clone 2-1#):

• Highly specific for a single epitope

• Consistent lot-to-lot performance

• IgG2b Kappa isotype with protein A+G affinity purification

• Recommended dilutions: 0.01-3μg/mL for WB; 5-30μg/mL for IHC/ICC

Polyclonal antibodies (e.g., 13970-1-AP):

• Recognize multiple epitopes

• Higher sensitivity but potentially lower specificity

• Rabbit IgG with antigen affinity purification

• Recommended dilutions: 1:500-1:2000 for WB; 1:100-1:400 for IHC

Selection between these antibody types should be guided by experimental requirements for sensitivity versus specificity and the specific application being used.

What emerging research areas involve HSPA1L detection and analysis?

Current research trajectories for HSPA1L include:

• Genetic association studies linking HSPA1L variants to preterm birth and other conditions

• Investigation of HSPA1L-vaspin complexes in kidney protection mechanisms

• Role of HSPA1L in cellular stress response and organelle dysfunction

• Potential therapeutic targeting of HSPA1L pathways in disease models

Researchers entering this field should consider interdisciplinary approaches combining genetic, proteomic, and functional studies to fully characterize HSPA1L's biological significance.

What methodological advances are improving HSPA1L research?

Recent technical advancements enhancing HSPA1L research include:

• Combined antibody detection with mRNA localization techniques to validate expression patterns

• In silico prediction tools for functional impact of genetic variants

• Improved co-localization methods for protein interaction studies

• High-throughput screening approaches for identifying HSPA1L modulators