C1orf50 Antibody

What is C1orf50 and what molecular characteristics should researchers know?

C1orf50 (Chromosome 1 Open Reading Frame 50) is an uncharacterized human protein encoded by the C1orf50 gene (ID: 79078) . Key characteristics important for antibody-based research include:

This discrepancy between calculated (22 kDa) and observed (26 kDa) molecular weight may indicate post-translational modifications and should be considered when verifying antibody specificity .

What types of C1orf50 antibodies are available and how do they differ in research applications?

C1orf50 antibodies are available in multiple formats with distinct research applications:

Monoclonal antibodies offer greater specificity and reproducibility for targeted epitopes, while polyclonal antibodies provide stronger signals through multiple epitope binding . For multicolor imaging studies, the conjugated versions eliminate cross-reactivity concerns from secondary antibodies .

What are the validated applications for C1orf50 antibodies and their optimal dilutions?

Different C1orf50 antibodies have been validated for specific applications with recommended dilution ranges:

For optimal results in immunohistochemistry applications, antigen retrieval with TE buffer pH 9.0 is suggested, though citrate buffer pH 6.0 may also be used as an alternative . Always titrate antibodies in each specific testing system to determine optimal working dilutions .

How should C1orf50 antibodies be stored and handled to maintain activity?

Proper storage and handling are essential for maintaining antibody activity:

• Storage temperature: Store at -20°C for long-term stability

• Format preservation: Most are provided in liquid format with stabilizers

• Buffer composition: Typically PBS with 0.02% sodium azide and 40-50% glycerol at pH 7.2-7.4

• Aliquoting: Recommended to avoid repeated freeze/thaw cycles, though some formulations are stable without aliquoting for -20°C storage

• Short-term storage: 4°C acceptable for brief periods (days to weeks)

• Shelf life: Typically 12 months when stored properly

The high glycerol content (40-50%) in most preparations prevents ice crystal formation during freezing and helps maintain antibody structure and function .

What factors should be considered when selecting a C1orf50 antibody for specific experimental applications?

When selecting a C1orf50 antibody, researchers should consider these critical factors:

• Epitope specificity: Different antibodies target distinct regions (e.g., AA 111-199)

• Species cross-reactivity: Most react with human samples, with varying cross-reactivity to mouse, dog, monkey, rat, and rabbit

• Application validation: Verify the antibody has been validated for your specific application (WB, IHC, IF, etc.)

• Immunogen considerations:

  • Full-length recombinant protein (NP_077002)

  • KLH-conjugated synthetic peptides

  • Fusion protein immunogens

• Detection method compatibility: Consider whether direct fluorophore conjugation or secondary antibody detection is preferable

• Clonality requirements: Monoclonal for high specificity and reproducibility, polyclonal for stronger signal

For studying potential protein interactions, particularly worth noting is that C1orf50 has been identified in SARS-CoV-2 protein interactome studies, which may influence antibody selection in infection-related research .

What controls and validation methods are recommended when working with C1orf50 antibodies?

Rigorous validation is essential for confidence in antibody-based experiments:

• Positive control samples:

  • HEK-293 cells have demonstrated positive detection

  • Human testis tissue shows reliable expression

  • Mouse kidney tissue provides a cross-species control

• Specificity validation methods:

  • Western blot size verification (expected band at 26 kDa)

  • Array validation against target protein + 383 non-specific proteins

  • Comparison against recombinant protein standards

• Negative controls:

  • Secondary antibody-only controls

  • Non-expressing tissue sections

  • Blocking peptide competition assays

• Antigen retrieval optimization:

  • Compare TE buffer pH 9.0 with citrate buffer pH 6.0

  • Test multiple dilutions in each system

For highest confidence results, orthogonal validation with multiple antibodies targeting different epitopes is recommended, particularly when studying an uncharacterized protein like C1orf50.

Why does the observed molecular weight of C1orf50 (26 kDa) differ from the calculated weight (22 kDa) in Western blotting?

The discrepancy between calculated (21.9-22 kDa) and observed (26 kDa) molecular weight of C1orf50 may result from:

• Post-translational modifications: Phosphorylation, glycosylation, or other modifications can alter migration in SDS-PAGE

• Structural characteristics: The protein's amino acid composition may affect SDS binding and migration

• Buffer conditions: Differences in sample preparation and buffer systems between experiments

• Protein sequence analysis: The full sequence (from aa 1-199) contains regions that may affect electrophoretic mobility

This discrepancy is consistently reported across sources and should be considered normal when validating C1orf50 antibodies . When troubleshooting Western blots, researchers should specifically look for bands at approximately 26 kDa, not the calculated 22 kDa.

What are common troubleshooting steps for inconsistent results with C1orf50 antibodies?

When facing inconsistent results with C1orf50 antibodies, consider these methodological adjustments:

• Western blot optimization:

  • Adjust antibody dilution within recommended range (1:500-1:2000)

  • Try extended blocking (5% BSA or milk for 1-2 hours)

  • Optimize primary antibody incubation time and temperature

  • Ensure fresh transfer buffers and complete protein transfer

• Immunohistochemistry improvements:

  • Compare antigen retrieval methods (TE buffer pH 9.0 vs. citrate buffer pH 6.0)

  • Test multiple dilutions (1:20-1:200 range)

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

  • Use amplification systems for weak signals

• Sample-specific considerations:

  • Different expression levels between cell/tissue types

  • Ensure target proteins maintain native conformation

  • Consider batch-to-batch variation in antibody production

• Signal detection optimization:

  • For fluorescent conjugates, minimize photobleaching and use appropriate filters

  • For enzymatic detection, optimize substrate incubation time

  • Consider signal amplification systems for low-abundance targets

How can C1orf50 antibodies be used in multi-protein detection experiments?

For simultaneous detection of C1orf50 with other proteins:

• Fluorophore selection for multiplexing:

  • AbBy Fluor® conjugated antibodies available in multiple wavelengths (350, 555, 647, 680, 750)

  • Select non-overlapping fluorophores to prevent bleed-through

• Strategic antibody pairing:

  • Combine rabbit polyclonal anti-C1orf50 with mouse monoclonal antibodies against other targets

  • For triple labeling, use directly conjugated antibodies to avoid species cross-reactivity

• Validation for co-localization studies:

  • Verify antibody specificity individually before multiplexing

  • Include appropriate single-stain controls

  • Use spectral unmixing for closely emitting fluorophores

• Application-specific considerations:

  • For flow cytometry: Titrate each antibody to determine optimal concentration

  • For IF: Consider sequential rather than simultaneous antibody application

  • For IHC: Test for potential epitope masking in multiple labeling

What is known about C1orf50's subcellular localization and protein interactions?

C1orf50 research is still emerging, with some key findings related to its localization and interactions:

• Subcellular localization:

  • Protein subcellular localization data is available through the Human Protein Atlas

  • Immunofluorescence detection with various C1orf50 antibodies enables localization studies

• Protein interactions:

  • C1orf50 has been identified in SARS-CoV-2 protein interactome studies

  • This suggests potential roles in viral-host interactions that warrant further investigation

• Methodological approaches for interaction studies:

  • Co-immunoprecipitation using monoclonal antibodies for high specificity

  • Proximity ligation assays for detecting protein interactions in situ

  • Use of recombinant C1orf50 protein (such as ABIN3090192) as controls or competitors