CBLB (Casitas B-lineage Lymphoma Proto-Oncogene B) is a gene that encodes an E3 ubiquitin-protein ligase involved in the regulation of immune cell activation and signaling.
It plays a key role in the negative regulation of antigen receptor signaling, thereby modulating immune responses.
The CBLB protein acts as a critical regulator of T-cell and B-cell function, and its dysregulation has been implicated in autoimmune disorders and cancer.
Researchers studying CBLB proteins can utilize PubCompare.ai, an AI-driven platform, to efficiently locate relevant protocols from scientific literature, preprints, and patents, and identify the best methods and products for their CBLB protein research.
This tool can enhance reproducibility, accuracy, and research effeciency in the field of CBLB protein studies.
Most cited protocols related to «CBLB protein, human»
Naive CD4+ T cells isolated from WT and Cblb−/− mice were stimulated with plate-bound anti-CD3 (2 μg/ml) and anti-CD28 (1 μg/ml) in the presence of Th1, Th2, Th17, or Th9 cytokine cocktails as previously described (Ying et al., 2010 ; Chang et al., 2010 ). Details are provided in the Supplemental Experimental Procedures.
Qiao G., Ying H., Zhao Y., Liang Y., Guo H., Shen H., Li Z., Solway J., Tao E., Chiang Y.J., Lipkowitz S., Penninger J.M., Langdon W.Y, & Zhang J. (2014). E3 Ubiquitin Ligase Cbl-b Suppresses Proallergic T Cell Development and Allergic Airway Inflammation. Cell reports, 6(4), 709-723.
Clinical validation of the LC-MS/MS assay was performed retrospectively using 79 DBS samples from 32 different patients with PA or MMACBL from the NBS and Metabolic Centers in Heidelberg (Germany), Szeged (Hungary) and Zurich (Switzerland). In 14 patients, the newborn specimens prior to therapy initiation were also available (PA: n = 4; CblB n = 1; CblC n = 9); all other samples were obtained during therapy. Storage duration of the patient specimens before second-tier analysis in median (range) was 0.61 (0.01-8.90) years. For additional characterization of samples, we used the first–tier acylcarnitine profiles (electrospray ionization-MS/MS [18 (link)–20 ]) determined both at time of blood sampling at the respective site providing the specimen, as well as after storage (measured at the NBS Centers Heidelberg and Szeged). First–tier cutoff values for C3 and C3/C2 in the respective laboratories were the following: Heidelberg 5.5 μM and 0.22; Szeged: 5.75 μM and 0.26; and Zurich: 6.0 μM and 0.17. The 3OHPA, MMA and MCA levels were also measured in External Quality Assurance DBS samples for NBS (n = 7) from ERNDIM. The validated assay is currently being evaluated prospectively in a pilot project for the extension of the German NBS panel named ‟Newborn screening 2020” at the NBS Center Heidelberg. In that prospective study, the reported LC-MS/MS assay is being applied as second-tier test using the same DBS specimen as in the primary screening. All procedures followed were in accordance with the ethical standards of the Helsinki Declaration of 1975, as revised in 2000. Written informed consent for regular newborn screening had been obtained from the parents of all individuals at time of blood sampling if required by national legislation. The anonymized, retrospective re-analysis of the specimens in the present study was approved by the Ethical Committees of the University of Szeged (217/2016-SZTE) and the University Children’s Hospital Zurich (2014-0211). At the University Hospital Heidelberg, parents of patients under treatment gave written informed consent for sample analysis for establishment of the second-tier strategies for the study NBS 2020. The study NBS 2020 was approved by the Ethical Committee of the University of Heidelberg (S-533/2015).
Monostori P., Klinke G., Richter S., Baráth Á., Fingerhut R., Baumgartner M.R., Kölker S., Hoffmann G.F., Gramer G, & Okun J.G. (2017). Simultaneous determination of 3-hydroxypropionic acid, methylmalonic acid and methylcitric acid in dried blood spots: Second-tier LC-MS/MS assay for newborn screening of propionic acidemia, methylmalonic acidemias and combined remethylation disorders. PLoS ONE, 12(9), e0184897.
Coronary artery specimens were obtained from autopsy from the Department of Pathology of the Amsterdam UMC and immediately fixed in 10% formalin and processed for paraffin embedding. All use of tissue was in agreement with the ‘Code for Proper Secondary Use of Human Tissue in the Netherlands’. CBL-B expression was analysed by immunohistochemistry, as described in the Supplementary material online. Gene expression of CBL-B in human atherosclerosis was examined by microarray-based transcriptional profiling of carotid endarterectomy specimens (BiKE dataset13 (link),14 (link)).
Seijkens T.T., Poels K., Meiler S., van Tiel C.M., Kusters P.J., Reiche M., Atzler D., Winkels H., Tjwa M., Poelman H., Slütter B., Kuiper J., Gijbels M., Kuivenhoven J.A., Matic L.P., Paulsson-Berne G., Hedin U., Hansson G.K., Nicolaes G.A., Daemen M.J., Weber C., Gerdes N., de Winther M.P, & Lutgens E. (2018). Deficiency of the T cell regulator Casitas B-cell lymphoma-B aggravates atherosclerosis by inducing CD8+ T cell-mediated macrophage death. European Heart Journal, 40(4), 372-382.
The 3′-UTRs of CBLB/GRB2 in Fig. 5a containing miR-27b putative target sites were amplified and cloned into psiCHECK-2 (Promega). A Fast Mutagenesis kit (VazymeBioTech) was used to mutate the miR-27b-binding sites of the CBLB and GRB2 3′-UTR vectors in Fig. 5a according to the manufacturer’s instructions. Dual-luciferase assays were performed using 1 × 104 Bads-200 cells per well in a 96-well plate. Following attachment for 8 h, the cells were co-transfected with 50 ng respective reporter constructs with either miR-27b or NC (50 nM). After 48 h, the Reporter Assay System Kit (Promega, 017319) was used to measure the luciferase activity. Each transfectant was assayed in triplicates. Firefly luciferase activity was normalized to constitutiverenilla luciferase activity.
Chen D., Si W., Shen J., Du C., Lou W., Bao C., Zheng H., Pan J., Zhong G., Xu L., Fu P, & Fan W. (2018). miR-27b-3p inhibits proliferation and potentially reverses multi-chemoresistance by targeting CBLB/GRB2 in breast cancer cells. Cell Death & Disease, 9(2), 188.
Total RNA and genomic DNA were extracted from patient PBMC or BM samples using the RNeasy and DNeasy Blood & Tissue Kits (Qiagen), respectively, following standard procedures, and concentrations were determined using Qubit® 3.0 Fluorometer. DNA targeted next-generation sequencing (NGS) analysis was performed using the TruSight Myeloid Sequencing panel (Illumina, San Diego, CA, USA) targeting 54 genes (full coding exons of 15 genes: BCOR, BCORL1, CDKN2A, CEBPA, CUX1, DNMT3A, ETV6/TEL, EZH2, KDM6A, IKZF1, PHF6, RAD21, RUNX1/AML1, STAG2, ZRSR2, and exonic hotspots of 39 genes: ABL1, ASXL1, ATRX, BRAF, CALR, CBL, CBLB, CBLC, CSF3R, FBXW7, FLT3, GATA1, GATA2, GNAS, HRAS, IDH1, IDH2, JAK2, JAK3, KIT, KRAS, KMT2A/MLL, MPL, MYD88, NOTCH1, NPM1, NRAS, PDGFRA, PTEN, PTPN11, SETBP1, SF3B1, SMC1A, SMC3, SRSF2, TET2, TP53, U2AF1, WT1). Amplicon sequencing libraries were prepared from 50 ng of DNA per sample. Input DNA quantitation was performed using a Qubit 3.0 Fluorometer with Qubit 1X dsDNA HS Assay Kit (Life Technologies). After quality control and equimolar pooling paired-end sequencing of the libraries was performed on a NextSeq (Illumina, San Diego, CA, USA) instrument with NextSeq 500 High Output v2 Kit to generate 2 × 150 read lengths according to manufacturer’s instructions. Sequence data were analyzed using the TruSeq Amplicon v3.0.0 app in BaseSpace™ Sequence Hub. After demultiplexing and FASTQ file generation, the software uses a custom banded Smith-Waterman aligner to align the reads against the human hg19 reference genome to create BAM files. Variant calling for the specified regions was performed using the Somatic Variant Caller (5% threshold, read stitching on).
Casado P., Rio-Machin A., Miettinen J.J., Bewicke-Copley F., Rouault-Pierre K., Krizsan S., Parsons A., Rajeeve V., Miraki-Moud F., Taussig D.C., Bödör C., Gribben J., Heckman C., Fitzgibbon J, & Cutillas P.R. (2023). Integrative phosphoproteomics defines two biologically distinct groups of KMT2A rearranged acute myeloid leukaemia with different drug response phenotypes. Signal Transduction and Targeted Therapy, 8, 80.
Alpha Thalassemia X-Linked Intellectual Disability Syndrome BCORL1 protein, human Biological Assay BLOOD BRAF protein, human calreticulin, human CBLB protein, human CBLC protein, human CDKN2A Gene CEBPA protein, human CSF3R protein, human Diploid Cell DNA, Double-Stranded Exons EZH2 protein, human FBXW7 protein, human FLT3 protein, human GATA1 protein, human GATA2 protein, human Genes Genome Genome, Human HRAS protein, human IDH2, human JAK3 protein, human Janus Kinase 2 K-ras Genes KDM6A protein, human MLL protein, human NPM1 protein, human NRAS protein, human Patients PTEN protein, human PTPN11 protein, human RUNX1 protein, human SETBP1 protein, human SMC3 protein, human Tissues TP53 protein, human ZRSR2 protein, human
Reagents were purchased from Merck-Life Sciences (Gillingham, UK) or Thermo Fisher Scientific (Loughborough, UK). Primary antibodies for: phosphor-AMPK (T172), AMPK, phospho-AKT (Ser473), AKT, phosphop44/42 (ERK1/2) (Thr202/Tyr204), ERK1/2,c-CBL, CAP, CBL-b, were from Cell Signaling (Leiden, NL); β-TUBULIN and ACTIN from Merck-Sigma-Aldrich; NDUFA from Thermo Fisher; CORE2, COX5A and SDHA from Abcam. HRP-conjugated secondary antibodies were from Thermo Fisher.
Aye C.C., Hammond D.E., Rodriguez-Cuenca S., Doherty M.K., Whitfield P.D., Phelan M.M., Yang C., Perez-Perez R., Li X., Diaz-Ramos A., Peddinti G., Oresic M., Vidal-Puig A., Zorzano A., Ugalde C, & Mora S. (2023). CBL/CAP Is Essential for Mitochondria Respiration Complex I Assembly and Bioenergetics Efficiency in Muscle Cells. International Journal of Molecular Sciences, 24(4), 3399.
Gene mutation analysis was performed using DNA extracted from bone marrow aspirate specimens in a small subset of patients. Amplicon-based next generation sequencing (NGS) targeting the entire coding regions of a panel of 81 genes was performed using a MiSeq platform (Illumina, San Diego, CA, USA) to detect somatic mutations and insertions and/or deletions as previously described [11 (link)]. The 81-gene panel included: ANKRD26, ASXL1, ASXL2, BCOR, BCORL1, BRAF, BRINP3, CALR, CBLB, CBLC, CBL, CRLF2, CREBBP, CEBPA, CSF3R, CUX1, DDX41, DNMT3A, EED, ELANE, ETNK1, ETV6, EZH2, FBXW7, FLT3, GATA1, GATA2, GFI1, GNAS, HNRNPK, HRAS, IDH1, IDH2, IKZF1, IL2RG, IL7R, KRAS, JAK2, JAK3, KDM6A, KIT, KMT2A, MAP2K1, MPL, NF1, NOTCH1, NPM1, NRAS, PAX5, PHF6, PIGA, PML, PRPF40B, PTEN, PTPN11, RAD21, RARA, RUNX1, SETBP1, SF1, SF3A1, SF3B1, SH2B3, SMC1A, SMC3, SRSF2, STAG1, STAG2, STAT3, STAT5A, STAT5B, SUZ12, TERC, TERT, TET2, TP53, U2AF1, U2AF2, WT1, and ZRSR2. All coding exons for each gene were covered with an analytical sensitivity of 5% mutant reads in a background of wild-type reads. Established bioinformatics pipelines were used to identify somatic variants.
Qiu L., Wang S.A., Tang G., Wang W., Lin P., Xu J., Yin C.C., Khanlari M., Medeiros L.J, & Li S. (2023). Blastoid B-Cell Neoplasms: Diagnostic Challenges and Solutions. Cancers, 15(3), 848.
ANKRD26 protein, human B Cell-Specific Transcription Factor BCORL1 protein, human Bone Marrow BRAF protein, human calreticulin, human CBLB protein, human CBLC protein, human CEBPA protein, human CRLF2 protein, human CSF3R protein, human Diploid Cell ETNK1 protein, human ETV6 protein, human Exons EZH2 protein, human FAM5C protein, human FBXW7 protein, human FLT3 protein, human GATA1 protein, human GATA2 protein, human Gene Deletion Genes HNRNPK protein, human Hypersensitivity IDH2, human IL2RG protein, human Insertion Mutation Interleukin 7 Receptor JAK2 protein, human JAK3 protein, human K-ras Genes KDM6A protein, human MAP2K1 protein, human MLL protein, human Mutation NPM1 protein, human NRAS protein, human Patients PRPF40B protein, human PTEN protein, human PTPN11 protein, human Renal Adysplasia RUNX1 protein, human SETBP1 protein, human SH2B3 protein, human SMC3 protein, human STAG1 protein, human STAT3 Protein STAT5A protein, human STAT5B protein, human telomerase RNA component TERT protein, human TP53 protein, human U2AF2 protein, human ZRSR2 protein, human
Nur77-GFP (Tg(Nr4a1-EGFP)GY139Gsat) transgenic mice, Zap-70 deficient mice lacking mature T cells (Zap70tm1Weis), and Foxp3-RFP mice (C57BL/6-Foxp3tm1Flv/J) have been previously described (Kadlecek et al., 1998 (link); Wan and Flavell, 2005 (link); Zikherman et al., 2012 (link)). C57BL/6J mice (WT mice in the text) and CD45.1 mice (B6.SJL-Ptprca Pepcb/BoyJ) were purchased from the Jackson Laboratory. A Nur77-GFP strain that is interbred with the OT-I (C57BL/6-Tg(TcraTcrb)1100Mjb/J) TCR transgenic strain was described previously (Au-Yeung et al., 2017 (link)). This OT-I-Nur77-GFP strain was interbred with a TCRα−/− strain (B6.129S2-Tcratm1Mom/J) purchased from the Jackson Laboratory. A Nur77-GFP strain interbred with the Foxp3-RFP strain has previously been described (Zinzow-Kramer et al., 2019 (link)). All mice were housed under specific pathogen-free conditions in the Division of Animal Resources at Emory University. Sts1−/−, and Cbl-b−/− strains were described previously (Carpino et al., 2004 (link); Chiang et al., 2000 (link)). These strains were maintained in the Laboratory Animal Resource Center at the University of California, San Francisco. Both female and male mice were used throughout the study. All animal experiments were conducted in compliance with the Institutional Animal Care and Use Committees at Emory University and the University of California, San Francisco.
Eggert J., Zinzow-Kramer W.M., Hu Y., Tsai Y.L., Weiss A., Salaita K., Scharer C.D, & Au-Yeung B.B. (2023). Accumulation of TCR signaling from self-antigens in naive CD8 T cells mitigates early responsiveness. bioRxiv.
Animals Animals, Laboratory Animals, Transgenic CBLB protein, human Females Institutional Animal Care and Use Committees Males Mice, Inbred C57BL Mice, Laboratory Mice, Transgenic NR4A1 protein, human Specific Pathogen Free Strains T-Lymphocyte UBASH3B protein, human ZAP70 protein, human
Serum-starved cells were grown to confluence before treatment with human HGF (catalog no.: CYT-244; ProSpec). Ligand concentrations and treatment times are indicated in figure legends. Cells were subject to two washes of PBS in room temperature followed by equilibration to 4 °C for 5 min. Cells were harvested in radioimmunoprecipitation assay buffer (150 mM sodium chloride, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, 10 mM sodium pyrophosphate, 100 mM sodium fluoride, 50 nM Tris [pH 8.0]) supplemented with protease inhibitor PMSF (2 mM; Calbiochem) and solubilized by end-over-end rotation for 10 min at 4 °C. Insoluble material was removed via centrifugation for 10 min at 4 °C. The resulting cell lysate was assessed by bicinchoninic acid assay (catalog no.: 23225; Thermo Scientific), diluted in SDS sample buffer, and boiled for 3 min prior to gel loading. Equal amounts of protein were loaded and resolved by 10% SDS-PAGE before transfer to nitrocellulose membrane. Membranes were immunoblotted using the indicated antibodies and the manufacturer’s instructions: total c-Met (catalog no.: 8198), phosphorylated c-Met pY1234/1235 (catalog no.: 3077), c-Cbl (catalog no.: 2747), Cbl-b (catalog no.: 9498), total p42/p44 MAPK (ERK1/2) (catalog no.: 4695) phosphorylated p42/p44 MAPK (ERK1/2) (catalog no.: 9101) (Cell Signaling Technologies), and α-tubulin (catalog no.: 6199) (MilliporeSigma). Following incubation with the appropriate horseradish peroxidase–conjugated secondary antibody (antimouse: catalog no.: 31450; Invitrogen; anti-rabbit: catalog no.: 7074; Cell Signaling Technologies), enhanced chemiluminescence was used to visualize immunoreactive bands in a Fotodyne imaging system. When comparing c-Met phosphorylation, samples were run on the same gel if possible (time courses, Figs. 2G and 3C). Otherwise, immunoblots were run, processed, and developed together (dose response, Fig. 3A). Each experiment was performed at least three independent times. Densitometry analysis was executed using the ImageJ software (National Institutes of Health). GraphPad/Prism 9 (GraphPad Software, Inc) was used for generating graphs and performing statistical analysis (57 , 62 (link), 75 (link)). Area under the curves were generated by plotting each individual experiment and finding the replicate areas before graphing in Prism.
Tarvestad-Laise K, & Ceresa B.P. (2023). Knockout of c-Cbl/Cbl-b slows c-Met trafficking resulting in enhanced signaling in corneal epithelial cells. The Journal of Biological Chemistry, 299(10), 105233.
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C57BL/6 mice are a widely used inbred mouse strain commonly used in biomedical research. They are known for their black coat color and are a popular model organism due to their well-characterized genetic and physiological traits.
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Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.
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Lipofectamine 3000 is a transfection reagent used for the efficient delivery of nucleic acids, such as plasmid DNA, siRNA, and mRNA, into a variety of mammalian cell types. It facilitates the entry of these molecules into the cells, enabling their expression or silencing.
Anti-Cbl-b is a research antibody product offered by Santa Cruz Biotechnology. It is directed against the Cbl-b protein, which is a member of the Cbl family of E3 ubiquitin-protein ligases. This antibody can be used in various research applications to study the Cbl-b protein and its role in cellular processes.
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Opti-MEM is a cell culture medium designed to support the growth and maintenance of a variety of cell lines. It is a serum-reduced formulation that helps to reduce the amount of serum required for cell culture, while still providing the necessary nutrients and growth factors for cell proliferation.
Cbl-b is a protein that functions as an E3 ubiquitin ligase, playing a role in the regulation of various cellular processes. It is involved in the ubiquitination and subsequent degradation of target proteins. The core function of Cbl-b is to facilitate the transfer of ubiquitin to substrate proteins, marking them for proteasomal degradation.
Rag1−/− mice are genetically engineered mice with a mutation in the Rag1 gene, which is essential for the development of mature T and B lymphocytes. These mice lack functional T and B cells, making them a useful model for studying immune system function and development.
The TruSight Myeloid Sequencing Panel is a targeted next-generation sequencing assay designed to detect genetic variants associated with myeloid neoplasms. The panel covers over 50 key genes related to myeloid malignancies, enabling comprehensive genetic profiling of samples.
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The Dual-Luciferase Reporter Assay System is a laboratory tool designed to measure and compare the activity of two different luciferase reporter genes simultaneously. The system provides a quantitative method for analyzing gene expression and regulation in transfected or transduced cells.
NSG mice are a strain of immunodeficient mice developed for use in biomedical research. They lack functional T cells, B cells, and natural killer cells, making them suitable for engraftment with human cells and tissues.
The CBLB (Casitas B-lineage Lymphoma Proto-Oncogene B) protein is an E3 ubiquitin-protein ligase that plays a crucial role in regulating immune cell activation and signaling. It acts as a negative regulator of antigen receptor signaling, helping to modulate immune responses. The CBLB protein is a critical regulator of T-cell and B-cell function, and its dysregulation has been implicated in autoimmune disorders and cancer.
Researchers studying CBLB proteins can utilize PubCompare.ai, an AI-driven platform, to efficiently locate relevant protocols from scientific literature, preprints, and patents, and identify the best methods and products for their CBLB protein research. This tool can enhance reproducibility, accuracy, and research effciency in the field of CBLB protein studies. PubCompare.ai allows you to screen protocol literature more efficiently, and leverage AI to pinpoint critical insights that help you identify the most effective protocols related to CBLB protein, human for your specific research goals. The platform's AI-driven analysis can highlight key differences in protocol effectiveness, enabling you to choose the best option for reproducibility and accuracy.
While the CBLB gene encodes a single protein, there can be different isoforms or variants of the CBLB protein due to alternative splicing or post-translational modifications. These variations may have slightly different structural or functional properties, which can impact their role in immune regulation and signaling pathways. Researchers should be aware of these potential CBLB protein variations when designing experiments and interpreting results.
