Recombinant PUS7L Antibody: Selection & Application Guide
Recombinant PUS7L Antibody: Selection & Application Guide
Recombinant PUS7L Antibody is a highly specific reagent designed to detect pseudouridine synthase 7-like (PUS7L) protein in a range of immunological applications, enabling researchers to investigate RNA modification pathways with precision and reproducibility.
- Product Name
- Recombinant PUS7L Antibody
- SKU
- Ab349233
- Antibody Type
- Recombinant monoclonal antibody
- Target Protein
- PUS7L (Pseudouridine Synthase 7-Like)
- Recommended Applications
- Western Blot (WB), Immunohistochemistry (IHC), Immunofluorescence (IF)
- Reference Clone / Source
- absea #RN-11168
Background: PUS7L Biology and Research Significance
PUS7L (Pseudouridine Synthase 7-Like) is a member of the pseudouridine synthase (PUS) enzyme family responsible for catalyzing the isomerization of uridine to pseudouridine (Ψ) in RNA substrates. Pseudouridylation is the most abundant post-transcriptional RNA modification in eukaryotic cells and plays critical roles in RNA structural stability, translation fidelity, and regulatory signaling pathways.
PUS7L has been implicated in the modification of transfer RNA (tRNA), ribosomal RNA (rRNA), and potentially messenger RNA (mRNA), making it a subject of increasing interest in epitranscriptomics research. Dysregulation of pseudouridine synthases, including PUS7L, has been associated with developmental disorders and certain disease states, underscoring the importance of reliable detection tools for this protein.
Recombinant antibodies targeting PUS7L, such as Ab349233, provide researchers with a reproducible, sequence-defined reagent to study PUS7L expression, localization, and function across diverse experimental models.
Antibody Specifications and Quality Attributes
The Recombinant PUS7L Antibody (Ab349233) is produced using recombinant technology based on the reference clone absea #RN-11168. Recombinant antibodies offer several key advantages over traditional hybridoma-derived antibodies:
- Sequence-defined: The antibody sequence is precisely known, eliminating batch-to-batch variability and ensuring long-term reproducibility across experiments.
- High specificity: Engineered to recognize the PUS7L target with minimal cross-reactivity to closely related pseudouridine synthase family members.
- Sustainable supply: Recombinant production removes dependence on live animals for ongoing antibody generation, supporting 3Rs principles in research.
Researchers should consult the specific product datasheet for Ab349233 for detailed information on host species, isotype, conjugation status, and validated reactivity profiles, as these parameters are critical for experimental design.
Recommended Protocols and Application Guidelines
The Recombinant PUS7L Antibody is suitable for use in several immunological detection techniques. The following general protocol guidance is provided to assist researchers in achieving optimal results:
Western Blot (WB)
- Prepare total cell or tissue lysate using an appropriate lysis buffer containing protease inhibitors.
- Resolve 20–40 µg of protein by SDS-PAGE and transfer to a PVDF or nitrocellulose membrane.
- Block the membrane with 5% non-fat milk or BSA in TBST for 1 hour at room temperature.
- Incubate with the primary Recombinant PUS7L Antibody at the manufacturer-recommended dilution overnight at 4 °C.
- Wash three times with TBST and incubate with an appropriate HRP-conjugated secondary antibody.
- Detect signal using enhanced chemiluminescence (ECL) substrate.
Immunohistochemistry (IHC)
- Deparaffinize and rehydrate FFPE tissue sections as appropriate.
- Perform antigen retrieval using citrate buffer (pH 6.0) or EDTA buffer (pH 9.0) as optimized for the specific tissue.
- Block endogenous peroxidase activity and apply protein block.
- Incubate with Recombinant PUS7L Antibody at the recommended dilution for 1 hour at room temperature or overnight at 4 °C.
- Apply appropriate secondary antibody and detection system; counterstain with hematoxylin.
Immunofluorescence (IF)
- Fix cells with 4% paraformaldehyde and permeabilize with 0.1–0.3% Triton X-100 if targeting an intracellular antigen.
- Block with 5% normal serum and incubate with primary antibody at recommended dilution.
- Detect with a fluorophore-conjugated secondary antibody and counterstain nuclei with DAPI.
Note: Optimal antibody dilutions and specific conditions must be determined empirically for each experimental system. Always refer to the Ab349233 product datasheet for validated dilution ranges.
Troubleshooting Common Issues
The following table outlines common issues encountered when working with recombinant antibodies such as Ab349233 and suggested corrective actions:
| Issue | Possible Cause | Suggested Action |
|---|---|---|
| No signal detected | Insufficient antigen expression; incorrect antibody dilution; failed antigen retrieval | Verify target expression in your cell/tissue model; optimize dilution; repeat antigen retrieval with alternative buffer |
| High background | Insufficient blocking; secondary antibody cross-reactivity; over-incubation | Extend blocking time; use species-matched controls; reduce primary antibody concentration |
| Multiple non-specific bands (WB) | Protease degradation; overloaded sample | Add fresh protease inhibitors; reduce protein load; optimize transfer conditions |
| Weak or inconsistent IHC staining | Sub-optimal antigen retrieval; tissue fixation variability | Titrate antigen retrieval conditions; standardize fixation protocol across samples |
If issues persist after following these guidelines, contact Aladdin Scientific technical support with your experimental conditions and lot number for personalized assistance.
Storage, Handling, and Quality Control
Proper storage and handling of the Recombinant PUS7L Antibody (Ab349233) are essential to maintain its performance over time:
- Storage temperature: Store at –20 °C for long-term storage. Avoid repeated freeze-thaw cycles, which can denature the antibody and reduce activity.
- Aliquoting: Upon first use, prepare single-use aliquots to minimize freeze-thaw cycles.
- Working solution: Diluted antibody solutions should be kept on ice during experiments and discarded after use; do not re-freeze diluted stocks.
- Stability: Refer to the specific product datasheet for the guaranteed stability period and expiration date.
- Quality control: Each lot of Ab349233 undergoes quality control testing to confirm target specificity and performance in validated applications. Lot-specific data are available on the product datasheet or upon request from Aladdin Scientific.
Always centrifuge the vial briefly before opening to collect the contents at the bottom, and handle the antibody under aseptic conditions to prevent microbial contamination.
Frequently asked questions
What is PUS7L and why is it an important research target?
PUS7L (Pseudouridine Synthase 7-Like) is an enzyme that catalyzes pseudouridylation, the conversion of uridine to pseudouridine in RNA molecules. This modification is critical for RNA stability and function. PUS7L is studied in the context of epitranscriptomics, RNA biology, and its potential links to developmental and disease-related phenotypes.
Which applications is the Recombinant PUS7L Antibody (Ab349233) validated for?
The Recombinant PUS7L Antibody (Ab349233) is recommended for use in Western Blot (WB), Immunohistochemistry (IHC), and Immunofluorescence (IF). Researchers should consult the product datasheet for specific validated dilution ranges and species reactivity, as optimal conditions may vary by application and sample type.
What are the advantages of using a recombinant antibody over a traditional hybridoma-derived antibody?
Recombinant antibodies offer several advantages: their amino acid sequences are precisely defined, eliminating batch-to-batch variability; they can be produced sustainably without continued animal immunization; and they can be engineered for specific properties such as isotype switching or conjugation. These features enhance reproducibility and reliability across experiments.
How should I store the Recombinant PUS7L Antibody to ensure maximum stability?
Store the Recombinant PUS7L Antibody at –20 °C for long-term use. To minimize degradation, aliquot the antibody into single-use volumes upon first receipt and avoid repeated freeze-thaw cycles. During experiments, keep working dilutions on ice and do not refreeze diluted antibody solutions.
What controls should I include when using the Recombinant PUS7L Antibody?
It is recommended to include a positive control using a cell line or tissue known to express PUS7L, a negative control omitting the primary antibody, and an isotype control matched to the antibody's host species and isotype. These controls help distinguish specific signal from background and validate experimental results.
References
- Schwartz S et al. Transcriptome-wide mapping reveals widespread dynamic-regulated pseudouridylation of ncRNA and mRNA. Cell. 2014.
- Safra M et al. The m1A landscape on cytosolic and mitochondrial mRNA at single-base resolution. Nature. 2017.
- Aladdin Scientific. Recombinant PUS7L Antibody (Ab349233) Product Datasheet. Aladdin Scientific.
Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.