lyrm1 Antibody

What is LYRM1 and why is it significant for research?

LYRM1 (LYR motif containing 1) is a novel nucleoprotein belonging to the mitochondrial leucine/tyrosine/arginine motif family of proteins. These proteins are characterized by short polypeptides containing an LYR motif near the N-terminus. LYRM1 has emerged as a significant research target due to its differential expression in metabolic tissues and its influence on cellular processes.

The significance of LYRM1 in research stems from several key observations:

• It shows highest expression in adipose tissue, with particularly elevated levels in obese subjects

• It is abundantly expressed in heart tissue, suggesting roles in cardiac function

• It promotes cell proliferation and inhibits apoptosis, affecting tissue homeostasis

• Overexpression causes mitochondrial dysfunction and is associated with insulin resistance

• It may serve as a mediator in obesity-related metabolic disorders

These characteristics position LYRM1 as a promising target for studies focused on obesity, metabolic disorders, and cardiovascular development, with potential implications for therapeutic interventions.

What is the cellular localization of LYRM1?

LYRM1 is primarily localized to the nucleus in cells, as confirmed through multiple experimental techniques. Anti-LYRM1 antibody binding has been visualized predominantly in nuclear fractions . This nuclear localization is consistent across different cell types and experimental systems, including:

• Adipocytes, where fusion protein studies with LYRM1-GFP constructs demonstrate nuclear accumulation

• Preadipocytes, where immunocytochemical detection shows nuclear localization

• Cell lines used for studying cardiac development, where nuclear staining patterns are observed

The product information for commercial LYRM1 antibodies consistently lists cellular localization as "Nucleus" . This nuclear distribution pattern is important for experimental design considerations when using LYRM1 antibodies, as it provides a reference pattern for validating antibody specificity and performance. The nuclear localization also informs hypotheses about LYRM1's functional role in gene regulation or nuclear signaling pathways.

What is the molecular weight of LYRM1 protein?

The calculated molecular weight of human LYRM1 is approximately 14 kDa, corresponding to its 122 amino acid sequence . This information is critical for Western blotting applications, where researchers need to identify the correct protein band.

"The actual band is not consistent with the expectation. Western blotting is a method for detecting a certain protein in a complex sample based on the specific binding of antigen and antibody. Different proteins can be divided into bands based on different mobility rates. The mobility is affected by many factors, which may cause the observed band size to be inconsistent with the expected size."

These discrepancies may result from:

• Post-translational modifications of the protein

• The presence of different modified forms simultaneously, resulting in multiple bands

• Alternative splicing of LYRM1, as referenced in product background information

When using LYRM1 antibodies for Western blotting, researchers should verify band identity through positive controls and consider additional validation approaches if band size differs from the expected 14 kDa.

How does LYRM1 influence cell proliferation and apoptosis?

LYRM1 exhibits dual regulatory effects on cell dynamics through distinct mechanisms affecting both proliferation and apoptosis. These functions have been extensively documented in both preadipocytes and cardiac cell models.

Proliferation enhancement:

Studies using MTT assays and cell cycle analysis revealed that LYRM1 dramatically increases cell proliferation rates . The specific effects include:

• Significantly higher growth rates in LYRM1-overexpressing cells compared to controls

• Increased percentage of cells in S-phase after serum stimulation

• This effect becomes apparent from 12 hours after serum stimulation

• Data from both assays consistently shows that LYRM1 promotes proliferation of preadipocytes

Apoptosis inhibition:

LYRM1 also demonstrates potent anti-apoptotic effects, as evidenced by:

• Reduced phosphatidylserine-positive cells (measured by annexin V-FITC binding) in LYRM1-overexpressing cells

• Decreased caspase-3 activity in cells with elevated LYRM1 expression

• Protection against serum deprivation-induced apoptosis

Together, these dual functions suggest LYRM1 plays a critical role in regulating cell population dynamics. By simultaneously promoting proliferation and inhibiting apoptosis, LYRM1 can significantly influence tissue homeostasis. In adipose tissue, this may contribute to expansion of the preadipocyte pool, potentially connecting LYRM1's elevated expression in obesity to adipose tissue growth .

What is the role of LYRM1 in heart development?

LYRM1 is abundantly expressed in heart tissue, suggesting potential roles in cardiac development. Research using P19 cells as a model system for cardiomyocyte differentiation has provided insights into its specific functions:

Differentiation effects:

• Overexpression of LYRM1 did not significantly affect the differentiation of P19 cells into cardiomyocytes

• No significant morphological differences were observed between LYRM1-overexpressing P19 cells and controls during the differentiation process

• Expression of cardiomyogenesis-specific markers (GATA4 and Nkx2-5) showed no difference between LYRM1-overexpressing cells and controls at the same time points

Cell dynamics regulation:

• LYRM1 dramatically increases proliferation of P19 cells

• LYRM1 protects P19 cells from apoptosis

• These effects suggest LYRM1 modulates the size of cardiac precursor cell populations rather than differentiation pathways

Based on these findings, researchers have concluded that "LYRM1 might have the potential to modulate cell growth, apoptosis, and heart development" . The gene may be particularly relevant in the context of congenital heart disease (CHD), as suggested by its expression pattern and functional characteristics . Future research may explore how alterations in LYRM1 expression or function contribute to cardiac developmental abnormalities.

What regulatory factors influence LYRM1 gene expression?

LYRM1 expression is regulated by multiple factors associated with metabolic health and insulin sensitivity. Understanding these regulatory mechanisms provides context for interpreting LYRM1's role in disease processes and suggests approaches for experimental manipulation of its expression.

Upregulating factors:

Downregulating factors:

This regulatory profile is particularly significant because it connects LYRM1 expression to factors known to modulate insulin sensitivity. The fact that insulin-sensitizing agents (rosiglitazone) decrease LYRM1 expression, while factors associated with insulin resistance (FFAs, TNF-α) increase its expression, supports the hypothesis that "LYRM1 may be an important mediator in the development of obesity-related insulin resistance" .

These findings provide important mechanistic insights for researchers investigating LYRM1's role in metabolic disorders and suggest experimental approaches for modulating its expression in cellular models.

What applications are LYRM1 antibodies validated for?

Commercial LYRM1 antibodies have been validated for several research applications, with varying levels of optimization for different techniques. The following table summarizes the validated applications across available antibodies:

When selecting a LYRM1 antibody for research, consider:

• Species reactivity requirements - many antibodies show cross-reactivity with human, mouse, and rat LYRM1

• Application-specific validation - choose antibodies tested in your application of interest

• Verified sample compatibility - preference for antibodies tested in tissues similar to your experimental system

• Detection method compatibility - ensure compatibility with your visualization system

Most commercial LYRM1 antibodies are rabbit polyclonal antibodies generated against recombinant human LYRM1 or fusion proteins containing LYRM1 sequences . This information is important when designing control experiments and interpreting cross-reactivity patterns.

How should LYRM1 antibodies be stored and handled?

Proper storage and handling of LYRM1 antibodies is crucial for maintaining their performance and extending their usable lifespan. Based on manufacturer recommendations across multiple suppliers, the following guidelines should be followed:

Storage conditions:

• Store at -20°C for long-term stability

• LYRM1 antibodies typically remain stable for 12 months when properly stored

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

Storage formulation:

• Most LYRM1 antibodies are supplied in buffers containing glycerol (typically 50%)

• Buffer formulations include PBS at pH 7.3-7.4

• Small amounts of preservatives like sodium azide (0.02-0.05%) are commonly added

Handling recommendations:

• Allow antibodies to warm to room temperature before opening to prevent condensation

• For antibodies without BSA, consider aliquoting into single-use volumes before freezing

• Some formulations (particularly smaller volume products) may contain BSA (0.1%) for added stability

• After use, return to -20°C promptly

Shipping considerations:

• LYRM1 antibodies are typically shipped with ice packs

• Upon receipt, transfer immediately to recommended storage conditions

Following these guidelines will help ensure consistent antibody performance across experiments and maximize the useful life of LYRM1 antibodies. Researchers should also consult product-specific documentation, as storage recommendations may vary slightly between manufacturers.

What controls should be included when using LYRM1 antibodies?

Robust experimental design requires appropriate controls to ensure reliable interpretation of results obtained with LYRM1 antibodies. The following controls should be considered:

Technical controls:

• Primary antibody omission control: Process samples following your normal protocol but omit the LYRM1 primary antibody to assess background from secondary antibody and detection system

• Isotype control: Use a non-specific antibody of the same isotype (typically rabbit IgG for most LYRM1 antibodies ) to assess non-specific binding

• Concentration-matched control: For more rigorous background assessment, use the same concentration of control antibody as your LYRM1 antibody

Biological controls:

• Positive tissue controls: Include tissues known to express high levels of LYRM1:

  • Adipose tissue (highest expression)

  • Heart tissue (high expression)

  • Verified samples listed by manufacturers (e.g., mouse adipose for WB, human liver cancer and tonsil for IHC)

• Negative tissue controls: Include tissues with minimal LYRM1 expression to assess non-specific background

• Expression-modulated controls: When possible, include:

  • LYRM1 overexpression samples (e.g., cells transfected with LYRM1 expression vector)

  • LYRM1 knockdown samples (using siRNA or similar approach)

Processing controls:

• Loading controls (for Western blot): Use housekeeping proteins (β-actin, GAPDH) to normalize LYRM1 signal

• Fixation controls (for IHC/IF): When working with fixed tissues, include samples prepared with different fixation protocols to assess effects on antibody binding

Implementing these controls will help distinguish specific LYRM1 signal from background or artifacts, leading to more reliable and interpretable experimental results.

How can I optimize immunostaining protocols for LYRM1 detection?

Optimizing immunostaining protocols for LYRM1 detection requires systematic adjustment of several parameters. Based on available technical information for LYRM1 antibodies, the following optimization strategy is recommended:

Antibody dilution optimization:

Begin with the manufacturer's recommended dilution range and test a series of dilutions to determine optimal signal-to-noise ratio. For LYRM1 antibodies, typical starting ranges are:

• IHC applications: 1:50-1:300

• Western blotting: 1:500-1:2000

Antigen retrieval optimization for IHC:

For formalin-fixed, paraffin-embedded tissues, test different antigen retrieval methods:

• Citrate buffer (pH 6.0)

• EDTA buffer (pH 8.0-9.0)

• Commercial retrieval solutions

• Compare different heating methods (microwave, pressure cooker)

Blocking optimization:

Test different blocking conditions to minimize background:

• Normal serum (5-10%) from the same species as the secondary antibody

• Bovine serum albumin (1-5%)

• Commercial blocking reagents

• Blocking duration (30 minutes to overnight)

Incubation parameters:

Optimize antibody incubation conditions:

• Duration: Compare standard incubation (1-2 hours at room temperature) vs. overnight at 4°C

• Temperature: Room temperature vs. 4°C

• Washing stringency: Test different wash buffer compositions and durations

Detection system selection:

Choose appropriate detection systems based on sensitivity requirements:

• For IHC: DAB-based vs. fluorescent detection

• For Western blot: Chemiluminescent vs. fluorescent detection

• Consider signal amplification systems for low-abundance detection

Sample preparation considerations:

• For Western blotting: Test different lysis buffers and protein extraction methods

• For IHC: Compare different fixatives and fixation durations

• For cell-based assays: Optimize cell density and attachment conditions

Document all optimization steps systematically, ideally in a grid format that allows comparison of multiple parameters simultaneously. After initial optimization, conduct reproducibility tests to ensure the protocol produces consistent results across experiments.

Why might observed molecular weight differ from the calculated weight for LYRM1?

The calculated molecular weight of LYRM1 is approximately 14 kDa (corresponding to its 122 amino acid sequence) , but researchers frequently observe discrepancies between calculated and detected molecular weights in Western blot applications. This phenomenon is acknowledged in antibody product documentation: "The actual band is not consistent with the expectation."

Several factors can explain these discrepancies:

Post-translational modifications:

• Phosphorylation: Addition of phosphate groups increases molecular weight and alters migration

• Glycosylation: Addition of carbohydrate moieties significantly increases apparent molecular weight

• Ubiquitination: This modification substantially increases observed molecular weight

Protein structural factors:

• Hydrophobic regions: Can bind more SDS and accelerate migration

• Charged domains: Can influence interaction with gel matrix and affect migration

• Incomplete denaturation: Residual structural elements can affect migration

Alternative splicing:

• LYRM1 gene undergoes alternative splicing resulting in multiple transcript variants

• Different isoforms may have different molecular weights

• Tissue-specific expression of different isoforms may occur

Technical considerations:

• Gel percentage: Different acrylamide percentages resolve proteins differently

• Running conditions: Buffer composition, temperature, and voltage affect migration

• Molecular weight standards: Calibration issues can affect size estimation

As noted in product documentation: "If a protein in a sample has different modified forms at the same time, multiple bands may be detected on the membrane."

When working with LYRM1 antibodies, researchers should:

• Include positive controls from tissues known to express LYRM1

• Consider the possibility of detecting multiple isoforms or modified forms

• Verify band identity through additional methods when critical for interpretation

• Document the apparent molecular weight observed in your experimental system

How do I interpret variable LYRM1 expression patterns across different tissues?

LYRM1 shows tissue-specific expression patterns that must be carefully interpreted in experimental contexts. Understanding this variability is essential for accurate data interpretation and experimental design.

Normal tissue expression pattern:

LYRM1 expression varies significantly across tissues, with a distinctive hierarchy:

• Highest expression: Adipose tissue, particularly omental adipose tissue

• High expression: Heart tissue

• Moderate expression: Liver and other tissues

• Wide distribution: LYRM1 is widely expressed across multiple tissues, but at varying levels

Pathological variation:

Expression levels change under certain conditions:

• Obesity: Significantly higher LYRM1 expression in adipose tissue of obese subjects compared to non-obese individuals

• Cancer: Detected in human liver cancer tissues (used as verified samples for antibody validation)

Cellular localization consistency:

Despite tissue-specific expression levels, cellular localization remains consistent:

• Nuclear localization is observed across different cell types

• This consistent localization pattern provides an important validation parameter for antibody specificity

Interpretation guidelines:

When interpreting LYRM1 staining patterns:

• Compare relative expression between tissues based on established patterns

  • Strong nuclear staining expected in adipocytes and cardiomyocytes

  • Weaker staining expected in other tissues

• Consider pathological context

  • Higher expression may be observed in tissues from obese subjects

  • Expression may be altered in disease states

• Validate unexpected patterns

  • Unusually high expression in typically low-expressing tissues requires validation

  • Altered subcellular localization (non-nuclear) should be critically evaluated

• Use quantitative methods when comparing expression levels

  • Western blotting with appropriate loading controls

  • qRT-PCR for mRNA level comparison

  • Image analysis of IHC with appropriate controls

Understanding these tissue-specific expression patterns provides important context for interpreting experimental results and can help identify potential technical issues versus genuine biological variations.

How can I troubleshoot weak or absent LYRM1 signal in my experiments?

When faced with weak or absent LYRM1 signal, a systematic troubleshooting approach can help identify and resolve the issue. Consider the following potential causes and solutions:

Antibody-related issues:

Sample-related issues:

Protocol optimization approaches:

• For Western blotting:

  • Increase protein loading amount

  • Extend transfer time for small proteins like LYRM1 (14 kDa)

  • Use PVDF membrane instead of nitrocellulose for better retention of small proteins

  • Extend primary antibody incubation (overnight at 4°C)

  • Increase sensitivity using enhanced detection reagents

• For IHC/ICC:

  • Optimize antigen retrieval (test different buffers and heating methods)

  • Reduce washing stringency

  • Use signal amplification systems (e.g., TSA, ABC method)

  • Block endogenous peroxidase activity (for HRP-based detection)

  • Extend antibody incubation times

• For all applications:

  • Test different blocking reagents to improve signal-to-noise ratio

  • Verify secondary antibody compatibility with primary

  • Include positive controls known to express LYRM1

If standard troubleshooting approaches fail, consider alternative detection methods or verification of LYRM1 expression at the mRNA level before revisiting protein detection.

What are the considerations for studying LYRM1 in the context of metabolic disorders?

LYRM1 has emerged as a significant factor in metabolic regulation, particularly in obesity and insulin resistance. Researchers studying LYRM1 in metabolic disorders should consider the following aspects:

Experimental model selection:

Different models offer distinct advantages for studying LYRM1 in metabolic contexts:

Key regulatory factors to assess:

When studying LYRM1 in metabolic contexts, consider measuring:

• Free fatty acid levels (upregulate LYRM1 expression)

• TNF-α (upregulates LYRM1)

• Effects of insulin-sensitizing drugs like rosiglitazone (downregulates LYRM1)

• Resistin levels (inhibits LYRM1 expression)

Functional readouts:

To link LYRM1 to metabolic outcomes, incorporate these assessments:

• Insulin-stimulated glucose uptake (reduced by LYRM1 overexpression)

• Mitochondrial morphology and function (abnormal with LYRM1 overexpression)

• Intracellular ATP synthesis (decreased with LYRM1 overexpression)

• Reactive oxygen species production (increased with LYRM1 overexpression)

Depot-specific considerations:

LYRM1 expression varies between adipose depots:

• Highest in omental adipose tissue of obese individuals

• Depot-specific differences should be accounted for in study design

• Different depots may show differential LYRM1 regulation

Translational perspectives:

When designing studies with clinical implications:

• Consider correlations between LYRM1 levels and metabolic parameters

• Evaluate LYRM1 expression before and after weight loss interventions

• Assess potential of LYRM1 as a biomarker for metabolic dysfunction

• Explore LYRM1 modulation as a potential therapeutic approach

This experimental framework will help researchers systematically investigate LYRM1's role in metabolic disorders, potentially leading to new insights into obesity-related pathophysiology and novel therapeutic targets.

What are emerging areas of investigation for LYRM1 research?

LYRM1 research is evolving, with several promising directions emerging from current findings. Researchers considering work in this field should be aware of these developing areas:

Mechanistic investigations:

• Elucidation of LYRM1's precise role in nuclear regulation and potential transcriptional effects

• Detailed characterization of how LYRM1 influences mitochondrial function and morphology

• Identification of LYRM1 interaction partners through proteomics approaches

• Determination of post-translational modifications affecting LYRM1 function

Pathophysiological relevance:

• Further exploration of LYRM1's contribution to obesity-related insulin resistance

• Investigation of LYRM1 in cardiovascular diseases, building on its established role in heart development

• Examination of potential roles in cancer biology, given its effects on cell proliferation and apoptosis

• Assessment of LYRM1 as a biomarker for metabolic dysfunction or cardiovascular risk

Therapeutic targeting approaches:

• Development of methods to modulate LYRM1 expression or activity

• Exploration of small molecule inhibitors targeting LYRM1 or its regulatory pathways

• Assessment of whether existing metabolic therapeutics affect LYRM1 expression or function

• Investigation of dietary or lifestyle interventions that normalize LYRM1 expression

Technological advances:

• Development of more specific and sensitive LYRM1 antibodies for research applications

• Creation of reporter systems for real-time monitoring of LYRM1 expression

• Application of gene editing approaches to create cellular and animal models with modified LYRM1 expression

• Integration of multi-omics approaches to understand LYRM1's role in complex metabolic networks

As LYRM1 research progresses, collaborative approaches combining expertise from metabolic research, cardiovascular biology, and molecular cell biology will likely yield the most comprehensive understanding of this multifunctional protein and its potential as a therapeutic target.