PTBP1 Antibodies

Polypyrimidine tract-binding protein 1 (PTBP1)

Activates exon skipping of its own pre-mRNA during muscle cell differentiation. Binds to the polypyrimidine tract of introns.

May promote RNA looping when bound to two different polypyrimidine tracts at the same pre-mRNA. May promote the binding of U2 snRNP to pre-mRNA.

Gene Information

Human
Gene Name:	PTBP1
Uniprot:	P26599
Entrez:	5725

Family

Belongs to:
No superfamily

Alternative Names

Heterogeneous nuclear ribonucleoprotein I; heterogeneous nuclear ribonucleoprotein polypeptide I; hnRNP I; HNRNPI; HNRNP-I; HNRPI; MGC10830; MGC8461; polypyrimidine tract binding protein (heterogeneous nuclear ribonucleoproteinI); polypyrimidine tract binding protein 1; polypyrimidine tract-binding protein 1; pPTB; PTB-1; PTB2; PTB3; PTB4; PTB57 kDa RNA-binding protein PPTB-1; PTB-T; RNA-binding protein

Molecular Weight

Mass (kDA):
57.221 kDA

Genomic Context

Human
Location:	19p13.3
Sequence:	19; NC_000019.10 (797452..812316)

Subcellular Localizations

Nucleus.

Diseases

• Nervousness
• Malignant Neoplasms
• Neoplasms
• Neuroblastoma
• Malignant Neoplasm Of Breast
• Glioma
• Adenocarcinoma
• Cell Transformation, Neoplastic
• Liver Carcinoma
• Glioblastoma
• Liver Diseases
• Malnutrition

• Insulinoma
• Myasthenic Syndromes, Congenital
• Myopathy
• Multiple Myeloma
• Mammary Neoplasms
• Stress, Psychological

Interacting Proteins

• PCBP1
• QKI
• RBM10
• SFPQ
• SNRPA

Pathways

• Translation
• Localization
• Secretion
• Cell Proliferation
• Insulin Secretion
• Rna Splicing
• Rna Processing
• Neurogenesis
• Mrna Processing
• Synapse Maturation
• Cell Activation
• Cell Migration
• Gene Silencing

• T Cell Activation
• Brain Development
• M Phase
• Oxidative Phosphorylation
• Nuclear Export
• Rna Surveillance
• System Development

PTMs

• Phosphorylation
• Cleavage

• Biotinylation
• Acetylation

For details, visit:
bosterbio.com/bosterbio-gene-info-cards/PTBP1

Boster Bio - Best Uses Of The PTBP1 Marker

The PTBP1 marker is a valuable tool to determine the binding partner for the lncRNA SNHG1. This guide will explain the qPCR primers that were used in the test, along with validation of kits for ELISA and RIP assays. It also provides practical tips for optimizing your experiments to assist you in optimizing your experiments. Boster Bio optimization guides also offer important information about experimental design and the results.

PTBP1 is a binder of lncRNA, SNHG1.

Hypoxia activates the LUCAT1 gene, which confers chemoresistance to tumor cells. It also physically interacts with PTBP1. This axis regulates alternative splicing of genes involved in DNA damage. Thus, PTBP1 could be a promising therapeutic target in hypoxic tumors.

To determine the SNHG1-PTBP1 lncRNA interaction Biotinylated RNA Pull-down test was conducted. Biotin-labeled, RNAs were made using T7 RNA Polymerase in vitro. Whole-cell lysates are incubated for 30 minutes at RT using RNAs labeled with biotin. Streptavidin magnetic beads were used to isolate the complexes of RNA-protein.

Whole-cell lysates were prepared using an EZMagna commercial RIP(tm) and a Millipore kit. The buffer contained magnetic beads and was incubated with the RIP buffer. These magnetic beads were used to conjugate primary antibodies to SNRP70and PTBP1 and other peptides. After incubation, the RNAs that were associated with peptides were analyzed using a quantitative real-time PCR.

PTBP1 functions as a binding partner for lncRNASNHG1. It protects p53 from interactions with cofactors that are involved in transcription-independent functions. PTBP1 is therefore a crucial regulator of SNHG1 transcription. However, it also inhibits the protein phosphorylation that is a retinoblastoma product.

BAFF is a new ligand for lncRNASNHG1. It is a ligand that binds SNHG1 as well as PTBP1. Together, they regulate the expression of lncRNAs that is SNHG1, a possible therapeutic target for DM. However, many studies are needed to confirm the findings.

PTBP1 regulates cytoskeleton remodeling in response to the microenvironment outside and inhibits the development of cancer. It is vital in the successful treatment of cancer due to its role in suppressing the growth of tumors and controlling the cytoskeleton's remodeling. This study provides a new approach to anticancer treatment.

QPCR primers that are used in the PTBP1 Marker

The PTBP1 Marker assay uses specific exon-exon junction primers. The targeted exons are identified as SON and hnRNP A2B1. Utilizing the PTBP2 qPCR assay, levels of RNA for PTBP2 were measured by measuring the ratio of PTBP1 intron 6-retained against total PTBP1.

The RT-PCR reaction used four microliters of dilute cDNA. Five ml of PerfeCta SYBR green FastMix, and 0.5 ml of reverse and forward primers. These reactions were performed in quadruplicate with MicroAmp Optical 384 well plates. The qPCR protocol consisted of a denaturation step at 95°C, 40 cycles at 95°C for 10 s and 30 cycles at 60°C for 30 s. After each cycle, melting curves were plotted in order to confirm that the PCR products had single peaks.

PTBP1 (a highly conserved RNA binding protein) is involved in the cell cycle, immune and other splicing steps. Furthermore, it aids in the migration of cancerous colorectal cells and regulates spermatogenesis as well as the proliferation of spermatogonia. Although its function in human development isn't yet fully known yet, it has been proved to be a key regulator of various cancer-related processes.

The knockdown of the PTBP1 gene was found to increase the level of mRNA of MCL1 in PC3 and h2299 cells. The knockdown of Flag-PTBP1 did not alter the levels of MCL1 transcripts in these cell lines. This suggests that the knockdown of PTBP1 affects the localization of MCL1 mRNA within the cell.

The PTBP1 gene contains 12 constitutive exons and two alternative exons. The 12 constitutive exons are identified by the boxed regions. The qPCR primers used in the PTBP1 Marker were constructed on these exons. The primers were validated with the malignant Glioma database.

RNA immunoprecipitation (RIP) assay

The use of RNA immunoprecipitation to trace the physical links between RNA molecules in vivo and proteins. RNA binding proteins are immunoprecipitated using specific antibodies, and the bound transcripts are identified using a variety of methods that include real-time PCR microarrays, and sequencing. This method was employed to study the physical relationship between the PTBP1 marker and the RNA in vivo.

To perform this assay the total proteins in RIPA buffer were separated by SDS-PAGE. The membranes were then blocked with dairy milk 5% defatted for 1 hour. Next, primary antibodies were incubated overnight at 4°C before being followed by secondary antibodies for the same duration. Anti-PTBP1 Cyclin D1, Cyclin D1, as well as Anti-b–actin were the main antibodies. The anti-p21 antibodies and anti-p16 ones were purchased from Abcam.

RIP assays for the PTBP1 marker also detect other cellular markers and transcripts. These genes can be assessed using RNA immunoprecipitation test (RIP). Primary neurons have been thoroughly tested for the PTBP1 marker. The expression level for any particular cell type is a reliable indicator of its health. RIP tests of PTBP1 can be particularly useful in ongoing research or in drug development.

The results of a recent RIP assay for the PTBP1 marker indicate that PTBP1 is a bona fide interconnected with LUCAT1. The RIP tests for LUCAT1 also reveal a connection between these two genes. CRC cells were used to conduct RIP tests of LUCAT1 (and PTBP1), which demonstrated that PTBP1's full-length was needed to form the strongest link with LUCAT1.

ELISA kit validation

Boster Bio spends a lot of time documenting a lot variability in order to ensure their PTBP1 ELISA kit is reliable and reproducible. The results are reliable and the coefficient of variation (CV) is usually less than 10%. Boster Bio ELISA Kit validation uses serum samples from healthy individuals to generate an dilution pattern. This lets you measure intra-assay or inter-assay precision.

Boster Bio's PTBP1 ELISA Kit has been validated against more than 250 samples. You can request the validation procedure and images are available upon request. Supervision polymer-based secondary antibodies are extremely efficient and can save the user up to 30 minutes of IHC or ELISA. Picoband is built on insights into immunogen design and is backed by technical support from BeNeLux distributor Sanbio.

Troubleshooting

The PTBP1 gene product, a ribosomal protein that regulates various aspects of the mRNA cycle, is a ribosomal product. These include translation, stability and localization, 3' end processing, translation and stability. It may also bind to DNA and act as a transcription factor. We will now discuss some of the most important aspects of PTBP1.

The expression of PTBP1 occurs during embryonic and postnatal development. This gene is essential in tumorigenesis, growth and other processes. It is involved in numerous processes, including the control of immune cells' activity. It is a fantastic candidate for the treatment of cancer and gene therapy. Its high expression can be used to identify other kinds of cancers and types.

Additionally, Ptbp1 plays a part in the noncanonical translation of the Per1 gene. In other words, it plays significant roles in the circadian rhythms of cells. PTBP1 regulates the translation of other circadian gene. It is unclear what PTBP1 regulates mRNA transcription. Whatever the reason, PTBP1 can be used to study circadian rhythms and pancreatic growth.