PPP1R13L Antibody, HRP conjugated
Description
Biological Context of PPP1R13L
PPP1R13L/IASPP is a conserved regulator of p53 and NF-κB pathways. It binds to p53 to inhibit apoptosis and interacts with NF-κB subunit p65/RelA to suppress transcriptional activity . Overexpression of PPP1R13L accelerates tumorigenesis in RAS/E1A-transformed cells, promoting metastasis while reducing p53 and active RelA levels . Its dual role as both a potential oncogene and tumor suppressor makes it a focal point in cancer research.
Key Studies Using PPP1R13L Antibodies
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Oncogenic Role: Overexpression of PPP1R13L in RAS/E1A-transformed mouse embryonic fibroblasts (MEFs) increased tumor formation and metastasis, correlating with reduced p53 stability .
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p53 Interaction: IASPP binds the ASPP2-binding domain of p53, competitively inhibiting ASPP2-mediated apoptosis .
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NF-κB Modulation: PPP1R13L suppresses NF-κB transcriptional activity by binding RelA/p65, influencing inflammatory responses .
Validation Data
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Western Blot: Detects bands at ~89 kDa (full-length IASPP) and smaller isoforms (e.g., 50 kDa) .
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Specificity: Validated using peptide blocking experiments, with immunogen sequences aligning across species (e.g., 100% homology in human, rabbit) .
Protocol Considerations
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Haplotypes composed of PPP1R13L rs1970764 and ATM rs11212592 may be linked to lung cancer. Haplotypes consisting of PPP1R13L, CD3EAP, and GLTSCR1 SNPs on Chr19q13.3 might be associated with lung cancer risk in the Chinese population. PMID: 30128886
- The expression of CD3EAP exon 1 was demonstrated to be significantly associated with PPP1R13L exon 1, while CD3EAP exon 3 was significantly associated with ERCC1 exon 11 in normal and non-small cell lung cancer (NSCLC) tissues. It was observed that short transcripts of ERCC1, CD3EAP, and PPP1R13L are co-expressed in the A549 NSCLC cell line. PMID: 29620255
- Increased expression of miR-150 suppressed the viability, proliferation, migration, and invasion of SW480 cells. Furthermore, iASPP was a direct target of miR-150 and played a key role in its anti-colorectal cancer (CRC) function. miR-150 may be a promising predictor of prognosis in CRC patients. PMID: 29750311
- IASPP knockdown suppressed cell viability and DNA synthesis capacity; the effect of miR-340 inhibition was partially attenuated by iASPP inhibition. PMID: 29982095
- The expression of iASPP was higher in high-grade astrocytic gliomas compared with low-grade astrocytic gliomas. PMID: 29257240
- Data indicated that iASPP could promote tumor growth by increasing autophagic flux, and iASPP could serve as a poor prognostic factor and a potential therapeutic target in lung cancer. PMID: 29072696
- Sertad1 could antagonize iASPP function by hindering its entrance into nuclei to interact with P53 in leukemic cells when iASPP was in the stage of overproduction. PMID: 29179704
- The interactive modulation among miR-124 and iASPP in p53-mutant or -deleted cells may serve as a crucial pathway, which mediates therapy resistance when p53 is mutated or deleted, in the process of photodynamic therapy treatment of Colorectal cancer. PMID: 29022915
- These findings indicated that XIST may regulate the tumor growth and metastasis via miR-140-dependent iASPP regulation. Taken together, our data indicated that XIST may be an oncogenic lncRNA that promotes the proliferation and metastasis of lung cancer through the regulation of miR-140 and could be regarded as a therapeutic target in human lung cancer. PMID: 28656261
- FHL2 and iASPP interacted with each other and co-localized in both nucleus and cytoplasm. Either FHL2 or iASPP silenced could reduce cell proliferation, induce cell cycle arrest at G0/G1 phase, and increase cell apoptosis. PMID: 28402264
- The restoration of miR-124 reduces iASPP expression and leads to p53-dependent tumor suppression, suggesting a therapeutic strategy to treat iASPP-associated cervical cancer. PMID: 27765948
- miR-124 regulates p63 via iASPP, while p63 targets miR-155 via the modulation of STAT1 expression in colorectal cancer. PMID: 28418858
- TP73-AS1 inhibited the brain glioma growth and metastasis as a competing endogenous RNA (ceRNA) through miR-124-dependent iASPP regulation. PMID: 29412778
- Findings revealed the detailed role of the miR-184/iASPP axis in Central nervous system lymphoma (CNSL) and this axis might modulate the proliferation and invasion of CNSL via regulating the PI3K/Akt signaling pathway. PMID: 28012196
- Data suggest that Keap1, rather than Nrf2, is critical for the recruitment of iASPP into the Keap1-Nrf2 complex PMID: 29033244
- Three htSNPs (haplotype-tagging single nucleotide polymorphism) (rs7354, rs14384, and rs3783501) covering 95% of the common haplotype diversity in 19p13.3-GADD45B and interaction of 19p13.3-GADD45B and 19q13.3-PPP1R13L and 19q13.3-CD3EAP variants and smoking-duration were explored in lung cancer risk Chinese. PMID: 28870783
- We describe the identification of a maternally inherited frameshift mutation in RAI1, causative for SMS. This is the first report about transmission of SMS from an affected parent to an offspring. PMID: 27683195
- These results determined PPP1R13L as the gene underlying a novel autosomal-recessive cardio-cutaneous syndrome in humans and strongly suggest that the fatal dilated cardiomyopathy during infancy is a consequence of failure to regulate transcriptional pathways necessary for tuning cardiac threshold response to common inflammatory stressors. PMID: 28069640
- UCA1 might promote proliferation and migration of glioma, to regulate the tumor growth and metastasis via miR-182 dependent iASPP regulation. PMID: 28137422
- Results demonstrate that iASPP is overexpressed in bladder cancer and promotes the malignancy of bladder cancer PMID: 28489738
- lncRNA H19 interacts with miR-140 to modulate glioma growth by targeting iASPP PMID: 27693036
- Increased expression of p53 and ASPP1 and downregulation of iASPP. PMID: 27177208
- Our study provides the first evidence that high iASPP-SV expression may be a novel prognostic factor and therapeutic target for glioma PMID: 26628298
- We were able to reproduce previously found associations between PPP1R13L and CD3EAP polymorphisms and lung cancer risk in an increased study group, and we found interactions between NFKB1 rs28362491-PPP1R13L rs1970764 and smoking duration and between CD3EAP rs735482 and smoking duration PMID: 26563375
- Inhibitor of apoptosis-stimulating protein of p53 (iASPP) was identified to be a direct target of miR-140 in pancreatic duct adenocarcinoma specimens and cell lines PMID: 26787707
- We show that iASPP is a novel substrate of caspases in response to apoptotic stimuli PMID: 26646590
- One novel region within PPP1R13L is hypomethylated in all transient neonatal diabetes type 1 patients included in this study PMID: 27075368
- Destabilization of p300/CBP by downregulation of iASPP expression levels appears to represent a molecular mechanism that contributes to chemoresistance in melanoma cells. PMID: 25675294
- One of the proteins identified, iASPP, showed reduced levels in the presence of GSK-3. Further, blocking iASPP activity increased cell death, particularly in p53 wild-type BC3 PEL cells. PMID: 26109723
- The present study showed that iASPP is overexpressed in oral cavity squamous cell carcinomas (OSCC) tissues and increased cytoplasmic iASPP is correlated with recurrence and poor survival outcomes in OSCC patients PMID: 25149434
- Results suggest that NFKB1 common variants and smoking duration and smoking duration-PPP1R13L rs1970764 interaction could be concerned with the lung cancer development in a Chinese population PMID: 25917613
- iASPP expression may act as a predictive marker of prostate cancer progression. PMID: 25341046
- Data highlight the importance of 14-3-3 proteins in antiviral responses and identify RelA-associated inhibitor and sirtuin 1 as novel regulators of antiviral innate immune responses. PMID: 24997996
- These data demonstrate that by interacting with desmoplakin and desmin, iASPP is an important regulator of desmosomal function both in vitro and in vivo. PMID: 25691752
- Authors have identified a novel mechanism modulating autophagy in keratinocytes that relies upon iASPP expression specifically reducing the interaction of Atg5-Atg12 with Atg16L1. PMID: 24777476
- MIRN124 binds to the 3'UTR of iASPP and suppressing mRNA expression and the proliferation of prostate tumor cells. PMID: 24966937
- Hematopoietic cells can be protected against apoptosis by iASPP. PMID: 24668753
- rs6966 (3' UTR of PPP1R13L, chr 19q13.32, P = 4.55 x 10(-9)) and rs414580 (intron 2 of MSR1, chr 8p22, P = 6.09 x 10(-8)) were significantly associated with ALL. PMID: 24604828
- Haplotype PPP1R13L rs4803817 polymorphism is associated with lung cancer risk. PMID: 24140460
- Overexpression of iASPP and the low expression of caspase-9 in esophageal cancer are closely correlated with tumor invasion and metastasis PMID: 24405603
- A higher rate of Helicobacter pylori infection, an increased expression of inhibitor of apoptosis stimulating protein of p53 (iASPP), and decreased expression of apoptosis-stimulating of p53 protein 2(ASPP2) was present in gastric cancer PMID: 23528480
- This study thus demonstrates that iASPP is highly elevated in human cervical cancer, and that overexpression of nuclear iASPP is correlated with poor prognosis and chemoresistance/radioresistance PMID: 23420450
- Downregulation of miR-124 promotes the growth and invasiveness of glioblastoma cells involving upregulation of PPP1R13L. PMID: 23624869
- The miR-124/iASPP axis can regulate the proliferation of colorectal cancer cells. PMID: 23691514
- PPP1R13L and CD3EAP variants may be associated with lung cancer risk in non-smoking Chinese women. PMID: 23624123
- Results suggest iASPP may contribute to the malignant progression of head and neck squamous cell carcinoma PMID: 22815155
- The PPP1R13L rs1970764 variant is a possible prognostic marker for patients with rectal cancer. PMID: 23180017
- When the Px(T)PxR motif is deleted or mutated via insertion of a phosphorylation site mimic (T311D), PP-1c fails to bind to all three ASPP proteins, ASPP1, ASPP2, and iASPP. PMID: 23088536
- These findings showed that iAPSS/iASPPsv reduced the growth inhibition and apoptosis induced by Dex or VP-16, with DNA damage accumulating which might promote the pathogenesis and/or progression of cancer. PMID: 22766503
- iASPP inhibited apoptosis independently of p53 in tumor cells, mainly by inhibiting the transcriptional activity of p63/p73 on the promoters of proapoptotic genes PMID: 22538442
Q&A
What is PPP1R13L and why is it an important research target?
PPP1R13L is a regulator that plays a central role in the regulation of apoptosis and transcription via its interaction with NF-kappa-B and p53/TP53 proteins. It blocks transcription of HIV-1 virus by inhibiting the action of both NF-kappa-B and SP1. Additionally, it inhibits p53/TP53 function, possibly by preventing the association between p53/TP53 and ASPP1 or ASPP2, therefore suppressing the subsequent activation of apoptosis . Recent research has shown that PPP1R13L is associated with poor prognosis in cervical cancer, making it a significant target for cancer research .
What applications are PPP1R13L antibodies commonly used for?
The HRP-conjugated PPP1R13L antibody is primarily validated for ELISA applications . Other non-HRP conjugated versions of PPP1R13L antibodies have been validated for Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), and Immunofluorescence (IF/ICC) . When selecting an antibody for your research, consider the specific application needs and whether the HRP conjugation is appropriate for your experimental design.
What is the optimal storage condition for PPP1R13L antibody, HRP conjugated?
Upon receipt, the antibody should be stored at -20°C or -80°C . It's crucial to avoid repeated freeze-thaw cycles as this can damage the antibody and reduce its effectiveness. For longer-term storage, aliquoting the antibody into smaller volumes is recommended to minimize freeze-thaw cycles .
How should I prepare samples for detection of PPP1R13L in different cell lines?
For optimal detection of PPP1R13L in cell lines such as SiHa, HeLa, MCF-7, and PC-3 (which have been validated for PPP1R13L expression), prepare cell lysates using standard lysis buffers containing protease inhibitors . When working with subcellular localization studies, it's important to note that PPP1R13L can be found in both cytoplasmic and nuclear fractions, with the full-length form (828 aa) detected in both locations while the N-terminal portion is entirely cytoplasmic . For nuclear extraction, use a specialized nuclear extraction protocol to ensure proper fractionation.
What positive controls should be included when using PPP1R13L antibody in research?
Based on validated reactivity data, positive controls for Western blot could include lysates from NIH/3T3, PC-3, MCF-7, HeLa (apoptosised), or C6 cells, as these have been confirmed to express detectable levels of PPP1R13L . For immunohistochemistry, human breast cancer tissue and human cervical squamous cancer tissue have been validated as positive controls .
How can I distinguish between different isoforms of PPP1R13L in my experiments?
PPP1R13L has different forms, including the full-length protein (828 aa) and cleaved fragments. The cleaved form PPP1R13L (295-828 aa) results from Caspase-3 processing and localizes primarily to the nucleus, while the full-length form is found in both cytoplasm and nucleus . When performing Western blot analysis, you should expect bands of approximately 100 kDa and 50 kDa in size, corresponding to isoforms 1 and 2 respectively . To specifically differentiate between these isoforms, consider using antibodies targeting distinct epitopes or combining with subcellular fractionation techniques.
What methodological approaches should be used to study PPP1R13L's interaction with p53 and p63?
To study PPP1R13L's interaction with p53 family proteins, researchers have utilized:
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Co-immunoprecipitation assays to demonstrate direct protein-protein interactions
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Luciferase reporter assays with P(site1)-luc and mutated P(△site1)-luc constructs to study transcriptional regulation
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RT-qPCR to assess the impact on target gene expression (e.g., PTEN mRNA expression levels)
Research has shown that PPP1R13L regulates PTEN transcription through its Ank-SH3 domain interaction with p53 family proteins. The C-terminal sequence variations in the Ank-SH3 domain lead to opposite functions compared to other ASPP family members .
How can I investigate PPP1R13L's role in cancer progression at the molecular level?
To investigate PPP1R13L's role in cancer progression:
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Perform knockdown and overexpression experiments using siRNA and expression vectors respectively
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Analyze effects on downstream signaling pathways such as PTEN/AKT/mTOR using Western blot
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Conduct RT-qPCR to quantify changes in target gene expression
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Use ChIP assays to identify direct binding sites on target gene promoters
Research has demonstrated that PPP1R13L knockdown significantly increased PTEN mRNA expression levels by 1.5 to 2-fold in cervical cancer cell lines, while overexpression reduced PTEN expression . This suggests PPP1R13L activates the PTEN/AKT pathway and influences the glycolytic phenotype in cancer cells.
What are common issues when using HRP-conjugated antibodies and how can they be addressed?
Common issues with HRP-conjugated antibodies include:
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High background: Increase blocking time/concentration, optimize antibody dilution, or add additional washing steps
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Weak signal: Ensure proper sample preparation, decrease antibody dilution, increase incubation time, or check substrate freshness
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Non-specific binding: Increase blocking, optimize antibody dilution, or pre-absorb with relevant tissues
For the PPP1R13L antibody specifically, the buffer (0.01M PBS, pH 7.4 with 0.03% Proclin 300 and 50% glycerol) is optimized for stability . If signal issues persist, consider testing the antibody with known positive controls such as human breast cancer tissue or PC-3 cell lysates.
How should I validate the specificity of the PPP1R13L antibody in my experimental system?
To validate antibody specificity:
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Include positive controls (PC-3, MCF-7, or HeLa cells) and negative controls
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Perform knockdown/knockout experiments using siRNA or CRISPR-Cas9 to verify signal reduction
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For advanced validation, use peptide competition assays with the immunogen peptide (83-102AA of Human RelA-associated inhibitor protein)
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Compare results using antibodies targeting different epitopes of PPP1R13L
When analyzing Western blot results, expect the full-length PPP1R13L to appear at approximately 110 kDa , though the calculated molecular weight is 89 kDa .
How can PPP1R13L antibodies be used to study its role in cervical cancer progression?
PPP1R13L is associated with poor prognosis in cervical cancer according to GSE52904 data . To study its role:
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Use IHC with the PPP1R13L antibody on cervical cancer tissue microarrays to correlate expression with clinical outcomes
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Compare PPP1R13L levels between normal cervical tissue and different stages of cervical cancer
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Investigate the molecular mechanism through which PPP1R13L suppresses p63 and regulates PTEN expression in cervical cancer cell lines
Research has shown that PPP1R13L drives cervical cancer progression by suppressing p63, and its knockdown significantly elevated PTEN mRNA expression levels . This suggests targeting PPP1R13L could be a potential therapeutic strategy.
What methodological considerations are important when investigating PPP1R13L's inhibition of HIV-1 transcription?
When studying PPP1R13L's role in HIV-1 transcription inhibition:
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Use reporter assays with HIV-1 LTR promoter constructs to measure transcriptional activity
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Perform ChIP assays to examine PPP1R13L's binding to the HIV-1 promoter region
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Investigate the interaction between PPP1R13L and both NF-kappa-B and SP1 transcription factors
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Use mutagenesis studies to identify critical domains of PPP1R13L involved in HIV-1 transcription inhibition
PPP1R13L blocks transcription of HIV-1 virus by inhibiting the action of both NF-kappa-B and SP1 , making it an interesting target for HIV research.
