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Publication Spotlight highlights the new and exciting discoveries citing Proteintech antibodies published in scientific journals across the globe.
What matters to Proteintech is the success of its antibodies in the hands of scientists - the people who make the discoveries that change and explain the world around us. In reaching 50,000 citations of Proteintech antibodies, our thanks go out to the talented researchers who made this milestone possible by using Proteintech antibodies in their work.
|Featured Articles - Click to navigate|
|Maintenance of CTCF- and Transcription Factor-Mediated Interactions from the Gametes to the Early Mouse Embryo.|
|Single-Cell Analysis of the Liver Epithelium Reveals Dynamic Heterogeneity and an Essential Role for YAP in Homeostasis and Regeneration.|
|Hypomorphic mutations of TRIP11 cause odontochondrodysplasia.|
|m6A enhances the phase separation potential of mRNA.|
|Meningeal lymphatic vessels at the skull base drain cerebrospinal fluid|
|The Polycomb Repressor Complex 1 Drives Double-Negative Prostate Cancer Metastasis by Coordinating Stemness and Immune Suppression|
|BCAA catabolism in brown fat controls energy homeostasis through SLC25A44|
|Protein kinase N controls a lysosomal lipid switch to facilitate nutrient signalling via mTORC1.|
|Fibroblast Growth Factor 21 Drives Dynamics of Local and Systemic Stress Responses in Mitochondrial Myopathy with mtDNA Deletions.|
|α-Ketoglutarate links p53 to cell fate during tumor suppression|
P53 is a famous tumor suppressor and is the most frequently mutated gene across a range of cancers. Although best known for its protective role in the DNA damage response, P53 has also recently been shown to affect cell metabolism. In a recent Nature paper (PMID: 31534224), Morris et al. showed that in pancreatic ductal adenocarcinoma (PDAC), P53 promotes a metabolic profile that inhibits tumor progression.
To determine the role of P53 in cancer metabolism, they modified a PDAC mouse model with a doxycycline-regulated shRNA targeting P53. With doxycycline treatment knocking down P53, tumors progressed rapidly to the malignant state; however, the subsequent restoration of P53 expression in the samples led to decreased tumor size and mortality. Comparing the tumor cells before and after doxycycline removal revealed a change in the metabolic profile characterized by an increase in the alpha-ketoglutarate (aKG) to succinate ratio, a key parameter controlling tumor fate.
In order to demonstrate that increased aKG was the major metabolic mediator of P53 in their model, they added aKG to PDAC cells directly and found it to have the same effects on gene expression and metabolism as restoring P53 expression. This was also observed when increasing aKG levels through the manipulation of Krebs cycle enzymes. Linking this metabolic shift to transcriptional reprogramming, Morris et al. then showed that aKG and P53 levels control the activity of aKG-dependent 5hmc epigenetic writers and that 5hmc levels are negatively correlated with the progression of disease. Thus, this work creates possibilities for novel metabolic approaches for treating PDAC and potentially other cancers that depend on P53 loss for progression.
Two Proteintech antibodies were used in this study: IDH1 (12332-1-AP) and OGDH (15212-1-AP). Both antibodies were utilized to confirm the efficiency of the doxycycline-regulated shRNA models used in their major experiments. 12332-1-AP has been validated in WB and ELISA, while 15212-1-AP has been validated in WB, ELISA, and IHC. The specificity of both antibodies has been demonstrated by KD/KO validation.
Catalog number: 15212-1-AP
Immunofluorescent analysis of 4% PFA fixed HeLa cells stained for anti-OGDH (15212-1-AP, 1:50, green) and anti-alpha tubulin (66031-1-IG, 1:100, red). DNA is stained by DAPI (blue).
About Scott Lowe, the Corresponding Author of the study:
Scott Lowe, PhD, is a professor at Memorial Sloan Kettering, chair of the Geoffrey Beene Cancer Research Center, and a Howard Hughes Medical Institute investigator. His work focuses on how the genetics of cancer cells enable tumorigenesis.
In a recent paper (PMID: 31523008) Forsström and colleagues examined the molecular causes of mitochondrial myopathies (MM), a class of neuromuscular diseases. The genetic cause of MM is mitochondrial DNA (mtDNA) defects. To better understand the etiology, Forsström et al. focused on the mitochondrial integrated stress response (ISRmt), a major metabolic cellular response to mtDNA mutations and alterations of mtDNA expression. In this study, they analyzed the effects of alterations of transcription factors, regulatory enzymes, and mechanistic target of rapamycin (mTOR) signaling in an in vivo mouse model and in biopsies of MM patients. They discovered that the early stages of the disease were characterized by increased levels of fibroblast growth factor 21 (FGF21) expression, which regulates ISRmt. Knocking out FGF21 resulted in a milder disease phenotype compared to controls in a MM mouse model. Elevated levels of FGF21 was shown to increase glucose uptake, serine de novo biosynthesis, and transsulfuration, which are characteristics of metabolic changes seen in MM, but it had no effect on the accumulation of mtDNA defects. The role of FGF21 depends on cell-type with the greatest impact on muscle cells and the dorsal hippocampus but little in fibroblasts. These findings raise the possibility of targeting FGF21 expression levels and function in individuals affected by MM.
Proteintech’s rabbit anti-MTHFD1L (16113-1-AP) was used in this study to examine the effects of FGF21 on the mitochondrial folate cycle. The authors determined that the expression of MTHFD1L (monofunctional C1-tetrahydrofolate synthase, mitochondrial), an enzyme converting glycine into formate, is regulated by FGF21. MTHFD1L antibody has been validated in Western blot, immunohistochemistry, ELISA, and immunoprecipitation, being specific for human, mouse, and rat.
Proteintech’s rabbit anti-CSE (12217-1-AP; Figure 1) was used in this study to examine transsulfuration in MM. The authors have shown that gamma cystathionase (CSE), an enzyme in the transsulfuration pathway, is elevated in MM patients and the MM mouse model and is regulated by FGF21. CSE antibody has been validated in Western blot, immunohistochemistry, ELISA, flow cytometry, immunofluorescence, and immunoprecipitation, being specific for human, mouse, rat, and rabbit.
Proteintech’s rabbit anti-CLPP (15698-1-AP) was used in this study as a marker of mitochondrial unfolded protein response (UPRmt). The authors have shown that caseinolytic mitochondrial matrix peptidase (CLPP), an early marker of UPRmt, is not part of ISRmt. CSE antibody has been validated in Western blot, immunohistochemistry, ELISA, immunofluorescence, and immunoprecipitation, being specific for human, mouse, rat, and zebrafish.
Catalog number: 12217-1-AP
Figure 1. Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using 12217-1-AP (Gamma cystathionase antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)
Mechanistic target of rapamycin (mTOR) signaling, mediated by mTORC1 and mTORC2 protein complexes, regulates a number of vital cellular processes. mTORC1 mainly acts as a nutrient sensor controlling rates of protein synthesis, cell growth, and autophagy.
In a recent Nature Cell Biology paper (PMID: 31451768), Wallroth and colleagues discovered that protein kinase N (PKN2), upon activation via mTORC2, positively regulates mTORC1 nutrient signaling by phosphorylating PI3KC2-β enzyme. PI3KC2-β is responsible for the synthesis of phosphatidylinositol- 3,4-bisphosphate at late endosomes and lysosomes, which then represses mTORC1 activity. However, PI3KC2-β, upon phosphorylation by PKN2, is sequestered in the cytoplasm by binding to 14-3-3 proteins, in turn preventing its inhibitory action on mTORC1 signaling. This signaling pathway was dissected using gain-of-function and loss-of-function cell models (engineered cell lines expressing tagged proteins, transient overexpression of protein mutants, depletion by siRNA, and studying KO cells) coupled with immunoblotting, immunoprecipitation, and immunofluorescence studies. This newly uncovered mechanism of active mTORC2’s influence on mTORC1 signaling represents a parallel route of mTORC1 activation by mitogens, which is classically attributed to AKT activation. Therefore, PKN2 may be an interesting drug target in diseases with altered nutrient signaling, e.g., in cancer or metabolic disorders – diabetes.
Proteintech’s rabbit anti-Rab35 antibody (11329-2-AP) was used in this study to examine the association of PI3KC2-β with various endosomal Rab GTPases. Rab35 is involved in the rapid recycling of receptors from early endosomes, as well as the transport of receptors from Rab11-positive recycling endosomes. Rab35 antibody has been validated in Western blot, immunohistochemistry, immunofluorescence, and immunoprecipitation, being specific for human, mouse, and rat forms of this protein.
Catalog number: 11329-2-AP
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 11329-2-AP (RAB35 antibody) at a dilution of 1:200 (under 40x lens) heat mediated antigen retrieved with Tris-EDTA buffer(pH9).
Warm-blooded, or endothermic, animals use a variety of mechanisms to maintain a constant core temperature. Brown adipose tissue (BAT) is the major organ involved in this process. To generate heat, BAT uncouples oxidative phosphorylation in the mitochondria from ATP generation, resulting in heat rather than stored energy. BAT is thought to exclusively use lipids and carbohydrates as fuel. However, writing in Nature, Yoneshiro et al. suggest that BAT also uses branched-chain amino acids (BCAAs) to generate heat under cold conditions and that inhibiting the use of this fuel source results in diabetes and glucose intolerance.
To gain insight into BAT function, Yoneshiro et al. first determined the metabolic differences between humans with high BAT activity. They found that individuals with high BAT activity have lower BCAA levels after cold exposure than those with low BAT activity. To test whether it is BAT that is responsible for breaking down BCAAs in cold conditions, Yoneshiro et al developed mice that lack the ability to catabolize BCAAs specifically in BAT. Consistent with this hypothesis, they found that BCAA levels do not change in these mice after cold stimulation. Additionally, disabling BCAA metabolism in BAT resulted in glucose intolerance and diet-induced obesity. Using a series of genetic and biochemical experiments, they showed that SLC25A44 is the key transporter of BCAAs into BAT mitochondria. Additionally, mice with knocked-down SLC25A44 have a lower core body temperature and cannot metabolize BCAAs after cold exposure. Overall, Yoneshiro et al.’s work uncovers a critical fuel source of BAT and may lead to insights into obesity and metabolic syndrome.
Proteintech’s anti-TOM20 antibody (11802-1-AP) was used as a mitochondrial marker to show that SLC25A44 is localized to this organelle and as a mitochondrial loading control. This antibody has been tested for WB, IP, IHC, IF, FC, and ELISA, in addition to KD/KO validation.
Catalog number : 11802-1-AP
Tested Applications: WB, IP, IHC, IF, FC, ELISA
Immunofluorescent analysis of ( 4% PFA ) fixed HepG2 cells using 11802-1-AP(TOM20 antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)
Prostate cancer is a leading cause of death in men worldwide. Despite the development of powerful immunotherapeutics, the immunosuppressive tumor microenvironment of prostate cancer metastases blunts the therapeutic response. In Cancer Cell, Su and colleagues show that hyperactivity of the polycomb repressor complex (PRC1) increases the expression of chemokines that attract Tregs and promote self-renewal of prostate cancer metastases (PMID 31327655).
Following on from previous work implicating PRC1 in prostate cancer tumorigenesis and metastasis, Su et al. began by comparing PRC1 levels in metastatic prostate cancer to other prostate cancer types and found that PRC1 is upregulated in metastatic samples. To test whether PRC1 is necessary for metastasis, they injected PC3 cells with knocked-down PRC1 into mice and discovered that metastatic lesions were decreased. Mechanistically, they found that PRC1 upregulation in cancer increases CCL2, which promotes stemness and recruits tumor-associated macrophages and Tregs to suppress the immune response. Demonstrating the potential clinical relevance of this result, the inhibition of PRC1 using a small molecule decreased metastasis and improved the efficacy of existing immunotherapeutics. Overall, Su et al.’s work opens the door to novel epigenetic approaches for combating metastasis, while PRC1 hyperactivity may be a common tumorigenic mechanism.
PRC1 is comprised of several subunits, among which RNF2 is critical to its activity. Proteintech’s anti-RNF2 antibody (16031-1-AP) was used to confirm the effectiveness of the investigators’ shRNA constructs targeting this protein and thereby disabling the complex. This antibody has been validated in WB, IP, IHC, IF, and ELISA.
Catalog number : 16031-1-AP
Tested Applications: WB, IP, IHC, IF, ELISA
human brain tissue were subjected to SDS PAGE followed by western blot with 16031-1-AP(RNF2 antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours
The clearance of macromolecules from cerebrospinal fluid (CSF) is crucial for the health of the brain. Impaired clearance may play a role in the pathogenesis of Alzheimer’s disease and other neurodegenerative diseases.
The current issue of Nature (PMID: 31341278) highlights the work of Ahn et al. on meningeal lymphatic vessels (mLVs) and their differential involvement in CSF drainage into cervical lymph nodes (cLN). Experimental investigation into mLVs up to this point has been based on dorsal mLVs, which are easily accessible but whose involvement in CSF drainage could not be proven. Gaining access to basal mLVs was more demanding due to their location at the skull base, close to an accumulation of blood vessels and cranial nerves. In this study, Ahn et al. found a method of overcoming these historical experimental limitations to the investigation of basal mLVs and compared the morphology and function of dorsal versus basal mLVs. Histological examinations of dorsal and basal mLVs revealed major differences. Basal mLVs exhibited an abundant number of branches that are similar to functional LVs and far better suited to the uptake of CSF than dorsal mLVs. Basal mLVs display properties of lymphatic pre-collectors due to their lack of SMCs, the mix of button and zipper-like endothelial cell junctions, and the occurrence of valves. In another approach, the authors tracked the lymphatic outflow of the CSF as well as the clearance of macromolecules from the CSF. Starting from the cisterna magna, the signal intensities of both, CSF contrast and macromolecular tracer, peaked in basal mLVs shortly after injection. In dorsal mLVs, the lymphatic outflow peaked relatively late, while uptake of the macromolecular tracer could not be detected, thus emphasizing the importance of basal over dorsal mLVs in CSF clearance.
Many neurodegenerative diseases are related to aging, where reduction of CSF clearance could be connected to mLV remodeling. Deletion of vascular endothelial growth factor 3 (VEGF3, a key player in mLV remodeling) in mice leads to regression of dorsal mLVs before basal mLVs are affected. No impairment of CSF clearance could be detected after regression of dorsal mLVs only; on the contrary, CSF clearance was reduced once VEGF3 deletion started to affect basal mLVs. These observations are an important starting point in tackling the connection between CSF clearance from brain-damaging macromolecules and many age-related pathologies.
Proteintech’s anti-PROX1 antibody (Cat# 11067-2-AP) was used as an immunofluorescence marker for the lymphatic endothelium in morphological studies comparing dorsal and basal mLVs. This antibody has been validated in Western Blot, immunofluorescence, flow cytometry, ChIP, and ELISA.
Catalog number : 11067-2-AP
Tested Applications: WB, IF, FC, CHIP, ELISA
Western blot image:
HepG2 cells were subjected to SDS PAGE followed by western blot with 11067-2-AP (PROX1 Antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.
Adapting for change is the key job of cells. In the last decade, researchers have discovered a new tool to dynamically influence the transcriptome to fine-tune cellular responses: methylated adenine or N6-methyladenosine (m6a). Advances in sequencing technologies have revealed that this epigenetic mark is abundant throughout the transcriptome and has been implicated in diverse processes including differentiation or the stress-response. The main question in the field is how this small chemical modification can have such powerful effects on the cell.
In a recent Nature paper (PMID: 31292544), Ries et al. show that m6a-RNA can be sequestered by their epigenetic readers into different phase-separated compartments during cell stress.
In an elegant set of biochemical experiments, the authors first showed that the m6a-RNA readers, YTHDF1-3, can reversibly separate into a different liquid phase from HEPES buffer by adjusting heat or salt. Afterwards, they found that heat shocks cause this protein to localize to stress granules in a separate liquid phase compartment with the aid of m6a-RNA. Using a METTL14 KO mutant that cannot catalyze m6a markings, they discovered that interaction between m6a and YTHDF1-3 is required for m6a-mRNA transcripts to be concentrated in stress granules during heat and chemical shocks. Ries et al.’s work suggests that m6a may be used to mark transcripts for protection during cellular stress and might be a mechanism for disease pathogenesis and tissue differentiation.
Proteintech’s YTHDF1 (17479-1-AP), YTHDF2 (24744-1-AP), and G3BP1 (13057-2-AP) antibodies were used in this study. 17479-1-AP and 24744-1-AP were used throughout the study to visualize YTHDF1-2 in mouse embryonic stem cells, U2OS, NIH3T3, and HEK293 cells. 13057-1-AP was used as a marker of stress granule formation. All three of these antibodies have been validated by KD/KO experiments to demonstrate specificity.
Catalog number: 17479-1-AP
Tested Applications: WB, IP, IF, IHC, ELISA
Western blot image:
The molecular causes of many human genetic diseases are yet to be identified. In a recent paper (PMID: 30728324), Wehrle and colleagues show that odontochondrodysplasia (ODCD), a form of skeletal dysplasia, is caused by hypomorphic mutations in the TRIP11 gene. TRIP11 encodes GMAP-210 (Golgi-associated microtubule-binding protein 210) which regulates protein transport and posttranslational modification within the Golgi apparatus. Severely reduced levels of GMAP-210 in patients’ cells disrupt the structure of the Golgi apparatus and abnormal glycosylation of cargo proteins, which negatively impacts chondrocyte differentiation.
The authors investigated posttranslational modifications of protein using western blotting and analyzing band patterns of glycosylated protein species and the resulting shifts of their molecular weights. Immunofluorescence imaging was used to look at the co-localization studies of Golgi proteins and IFT20, a binding partner of GMAP-210 that localizes both to the Golgi and primary cilium.
Proteintech’s rabbit anti-IFT20 antibody (13615-1-AP) was used in this study to show that GMAP-210 protein mediates correct subcellular localization of IFT20 to the Golgi apparatus. This antibody has been validated in western blot, immunohistochemistry, and chromatin immunoprecipitation with genetic knockdown data to demonstrate its specificity.
Catalog number: 13615-1-AP
Tested Applications: WB, IP, IHC, IF, ELISA
IF image: Immunofluorescent analysis of (10% Formaldehyde) fixed MDCK cells using 13615-1-AP (IFT20 antibody) at dilution of 1:100 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)
Reviews: 5 star
Verified customer: "Excellent for IF labelling of the Golgi under PFA fixation."
For more cilia-related antibodies, please see our cilia card.
Additionally, Proteintech’s rabbit anti-decorin antibody (14667-1-AP) was used to look at posttranslational modifications of extracellular matrix components.
The liver plays a crucial role in protein, carbohydrate, and lipid metabolism, the detoxification of drugs, and the breakdown of hormones. Two cell types, hepatocytes and biliary epithelial cells (BECs), are critical to the regenerating capabilities of the liver.
The current issue of Cell Stem Cell (Vol. 25, issue 1) features the work of Pepe-Mooney and coworkers who elucidated the dual role of the YAP signaling pathway in liver cell homeostasis and the initiation of regenerative processes (PMID: 31080134). YAP is a transcription factor that is involved in the regulation of cell proliferation as well as apoptosis. In an elegant assay, the authors utilized FACS followed by a single-cell RNA-seq to compare the expression profiles of 2,344 homeostatic BECs. Upon the differential expression of several YAP target genes, they identified two major cell clusters, YAP activated and non-activated cells. To identify the reason for this heterogeneity in YAP expression, the assay was repeated for BECs and hepatocytes after drug-induced mimicking of chronic liver damage in vivo. The results revealed the importance of YAP expression in the regenerative response of BECs and hepatocytes under liver-damaging conditions. The exposure of BECs to physiological levels of bile acid (BA) revealed the additional role of the YAP signaling pathway in cell homeostasis, dependent on intracellular BA uptake. This study presents a great basis for further investigations to elucidate cell type-dependent roles and the complexity of YAP signaling.
Beside the two major clusters, the initial RNA-seq screening revealed a small subset of cells co-expressing Dmbt1 and Ly6d, which are markers for extrahepatic BECs.
Proteintech’s Rabbit anti-LY6D (Cat# 17361-1-AP) was used for immunofluorescence staining as a differential marker for extrahepatic BECs. Based on the antibody performance it was possible to separate extrahepatic from intrahepatic BECs. This antibody has additionally been validated in Western Blot, immunohistochemistry, and ELISA.
Catalog number: 17361-1-AP
Tested Applications: WB, IHC, ELISA
Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 17361-1-AP( LY6D Antibody) at dilution of 1:50 (under 40x lens)
Is a child being born to a home with food insecurity? A frigid climate? A place with high toxic exposure? Being able to endow children with genetic adaptations to the environment provides significant evolutionary fitness. The prevailing notion is that these tweaks in the gamete stage are mediated by methylation and histone modifications. In the cover story for July’s Molecular Cell (PMID: 31056445), Jung et al. suggest that transcription factor sites and enhancer loops are also inherited from the parents.
Using a transposon accessibility assay and ChIP-seq, they show that mature sperm have transcription machinery and enhancer loops at many sites, but that the corresponding genes are not actively transcribed until embryogenesis. Comparing the loops of DNA in the parental gametes and embryos, they further showed that many of the CCCTC-binding factor (CTCF) and other transcription factor loops are inherited in a parent-specific manner. Their work introduces chromatin structure and early transcription factor priming as novel parental imprinting mechanisms for further investigation.
Proteintech’s Znf143 antibody (16618-1-AP) was used in this study to show that Zinc-finger transcription factors are present at many of inherited sites. This antibody has been validated in Western blot, immunohistochemistry and chromatin immunoprecipitation with genetic knockdown data to demonstrate its specificity.
Catalog number: 16618-1-AP
Tested Applications: WB, IHC, ChIP
Western blot image:
Mouse brain tissue were subjected SDS PAGE followed by Western blot with 16618-1-AP (Znf143 antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.