The encoded protein is a major constituent of the contractile apparatus and one of the two nonmuscle cytoskeletal actins that are ubiquitously expressed. The effect of bowel inflammation on HKG remains unknown. Relevance to Autism. endogenous reference (or ‘housekeeping’) gene for normalization of data. Results: Evaluation of the expression levels of the 6 target genes showed that ACTB, GAPDH, and RPL13A show higher expression compared to SDHA, POLR2A, and TBP. Actins are highly conserved proteins that are involved in cell motility, structure and integrity. ACTB is generally regarded as a constitutive housekeeping gene with the assumption that ACTB expression is usually unaffected by most experimental or physiological conditions and it has therefore been widely used as a reference for quantification of changes of genes/proteins in cells and tissues [6], [7], [8]. Essential component of cell cytoskeleton; plays an important role in cytoplasmic streaming, cell shape determination, cell division, organelle movement and extension growth. Each of these four HKGs is represented by 50 spots in the array. Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells. Includes 10 individual primer sets for Actb, B2m, … Cancer development affects almost all pathways and genes [1–4].Also affected are the so-called "housekeeping" genes, which are involved in the cell's common basic functions [5–8].Typical housekeeping genes include glyceraldehyde-3-phosphate dehydrogenase (GAPDH), beta-actin (ACTB), TATA-binding protein (TBP), ribosomal proteins (RP), and many more [9–14]. Gene ID: 396526, updated on 6-May-2021. Beta-actin (human gene and protein abbreviation ACTB/ACTB) is one of six different actin isoforms which have been identified in humans. The encoded protein is a major constituent of the contractile apparatus and one of the two nonmuscle cytoskeletal actins that are ubiquitously expressed. For example, housekeeping genes such as actin, ubiquitin or ribosomal genes are thought to … Beta-actin (ACTB) has traditionally been regarded as an endogenous housekeeping gene and has been widely used as a reference gene/protein in quantifying expression levels in tumors. Version: 14-September-2017 Alternate Names: Gene Info: Publications: Filby et al., 2007, Appropriate 'housekeeping' genes for use in expression profiling the effects of environmental estrogens in fish., BMC Mol. This gene encodes one of six different actin proteins. In a study of human asthma, where target gene expression between experimental groups appeared different when ACTB was used as a housekeeping gene, the results were misleading because it was in fact the ACTB, rather than the target gene, that changed 31. HKGs for potential use, namely ACTB, GAPDH, RPL5, and LDHA. Taken together, our data show the relevance of previous validation of candidate control genes for each experimental design and indicate that the most suitable housekeeping genes are SDHA and ACTB for quantitative PCR analysis in glioma tissue of different malignancies, after recurrence, and under different treatment conditions. Normalize real time PCR results to multiple housekeepers rather than one for more accurate quantitation. SUMMARY OF HKGS AND TARGET GENE USED IN THIS STUDY. ACTB is generally regarded as a constitutive housekeeping gene with the assumption that ACTB expression is usually unaffected by most experimental or physiological conditions and it has therefore been widely used as a reference for quantification of changes of genes/proteins in … ( b ) Copy number transcript by well of PSMB6 , PGGT1B , UBQLN2 and UQCR2 . GeneRIFs: Gene References Into Functions. Instability of housekeeping genes (HKG), supposedly unregulated and hence used as normalizers, may dramatically change conclusions of quantitative PCR experiments. Alpha actins are a major constituent of the contractile apparatus. Actins are highly conserved proteins that are involved in cell motility, structure, integrity, and intercellular signaling. Includes 8 individual primer sets for Actb, B2m, Gapd, Gusb, Hprt1, Pgk, Pp1a, and Rpl13a. Symbol Gene name Function Housekeeping genes GAPDH Glyceraldehyde-3-phosphate dehydrogenase Glycolytic enzyme An endogenous control gene is a gene whose expression level should not differ between samples, such as a housekeeping or maintenance gene. Flowchart of a novel identification strategy for housekeeping genes (nHKGs) in breast cancer. The genes encoding β-actin (ACTB in human or Actb in mouse) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH in human or Gapdh in mouse) are the two most commonly used references for sample normalization in determination of the mRNA level of interested genes by reverse transcription (RT) and ensuing polymerase chain reactions (PCR). However, ACTB is closely associated with a variety of cancers and accumulating evidence indicates that ACTB is de-regulated in liver, melanoma, renal, colorectal, gastric, pancreatic, esophageal, lung, breast, prostate, … The maximum and minimum expression ranges were 22.68 cycles for GUSB and 8.61 cycles for ACTB respectively (Table3). 0’s proportion is defined as the proportion of different cell lines in which a given gene is not expressed; and CV is the ratio of the standard variation to the mean. Statistical significance was evaluated by paired Student’s t-test using the GraphPad Prism software. Product Description. Perform up to 1000 assays (based on 20 ul assay volume) for each housekeeping gene. Another example comes from a breast cancer housekeeping validation study, where PUM1 (Pumilio Homolog 1) was a better option as the internal control in this type of cancer, where B2M (β-2-microglobulin) and ACTB showed significant differences ), ribosomal genes (multiple genes for … We selected genes which were recently described as very stable (RPL27, RPS29, RPS13, OAZ1) and two commonly used housekeeping genes (GAPDH, ACTB). A list of 7780 genes was first obtained by selecting genes with low 0’s proportion and coefficient of variation (CV). summary, ACTB and SDHA exhibited the best stability values and showed the lowest intergroup expression variability. To refine our ECFC housekeeping gene selection and establish the ideal number of housekeeping genes to be used in a panel, we designed and successfully validated RT-qPCR primers for 28 target genes , including targets from geNorm screening of microarray data plus “classical” housekeeping genes i.e., ACTB, GAPDH, and UBC. Normalized signal intensities were compared TABLE 1. Typical housekeeping genes relate to the basic functioning of cells, such as cytoskeletal genes (Actb (actin), Tubb5 (tubulin), Vim (vimentin) etc. housekeeping genes, including ACTB (ß-Actin) and GAPDH4. This is one of the two nonmuscle cytoskeletal actins. Proteins in this family are organized into a network of fibers called the actin cytoskeleton, which makes up the structural framework inside cells. Summary Other designations. We are the first to report a novel panel of species-specific housekeeping genes (HKG) for CHO cells. Summary This gene encodes one of six different actin proteins. 16 An additional member of this group of ribosomal protein-encoding genes, which may also be suitable for housekeeping … Normalize real time PCR results to multiple housekeepers rather than one for more accurate quantitation. ACTB Ensembl Gene Name Gene Info: Gene Biotype PROTEIN_CODING Publications: NCI: ACTB. A de novo missense variant in the ACTB gene has been identified in an ASD proband from the Autism Sequencing Consortium (De Rubeis et al., 2014). At present, so-called housekeeping genes are universally used as a reference 13. The range of threshold cycles (CT Range) was between 8.61 and 22.68 among endogenous candidate genes with a mean value ranging from 25.1 (± 0.5 s.e.m) for ACTB to 32.61 (± 0.43 s.e.m) for GUSB. Commonly used housekeeping genes in Q-RT-PCR include beta actin (ACTB), glyceraldeyde-3-phosphate dehydrogenase (GAPDH), ribosome small subunit (18S) ribsosomal RNA (rRNA), Ubiquitin C (UBC), hypoxanthine guanine phosphoribosyl transferase (HPRT), succinate dehydrogenase complex, subunit A (SDHA) and Tyrosine 3-monooxygenase/tryptophan 5 … A subset of housekeeping genes, including 18S, ACTB, CYPA, GAPDH, PGK1, UBC, UBXN11 and YWHAZ, exhibited variable Cq values. Each bar represents the mean of triplicate measurements on three different samples for each cell line. Genes with r 2 closer to 1 are considered as the most stable genes; thus ywhaz and alas1 with r 2 = 0.996 followed by actb (r 2 = 0.978) are ranked as the highest stable genes, while ubc (r 2 = 0.835) has the lowest value, indicating it had the lowest stability expression within these selected candidate reference genes as shown in Table 3. ACTB, a housekeeping gene in humans, mice, rats and other experimental animals, is widely used as an internal reference in quantitative methods, including qPCR and western blot analysis, which are two popularly methods used for the quantification of the mRNA and protein content, respectively. Established housekeeping genes in meningioma RT-qPCR experiments are genes such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and β-Actin (ACTB) [ 10 – 18] as well as ribosomal RNA (18S rRNA) and TATA binding box protein (TBP) [ 19 – 21 ]. Actins are highly conserved proteins that are involved in cell motility, structure, integrity, and intercellular signaling. actin, cytoplasmic 1, Bact; actin, beta-actin, cytoplasmic 1. of the most used housekeeping genes, was not a good choice for this carcinoma 17. Reference genes, frequently termed “housekeeping genes,” are used to normalize the expression results for differences in cDNA quantity between different specimens and thus enable comparisons between genes of interest across treatments. Here, we have analysed the expression stability of 12 commonly used reference genes (Actb, B2m, Gapdh, Hprt, Pgk1, Rn18s, Rpl13a, An ideal endogenous control generally demonstrates gene expression that is relatively constant and highly abundant across tissues and cell types. Among its related pathways are G-Beta Gamma Signaling and Apoptosis Modulation and Signaling . Gene Ontology (GO) annotations related to this gene include identical protein binding and RNA polymerase II proximal promoter sequence-specific DNA binding . An important paralog of this gene is ACTG1. Results show that despite being commonly used housekeeping genes neither GAPDH mRNA nor beta-actin mRNA is expressed in all cell types or tissues at high levels. PROX1 and β -actin mRNA levels were quantified and PROX1 expression normalized against the expression of the ACTB housekeeping gene. Product Description. These genes were selected based on expression stability from data collected from more than 25 experimental conditions in various CHO cell lines using our custom microarray platform. The ACTB gene provides instructions for making a protein called beta (β)-actin, which is part of the actin protein family. Thus, they are expressed in all cells of an organism under normal and patho-physiological conditions, irrespective of tissue type, developmental stage, cell cycle state, or external … Entrez Gene Summary for ACTB Gene. Biochemical studies on transcription initiation of the housekeeping gene promoters have been difficult, partly due to the less-characterized promoter motifs and transcription initiation process. Numerous pseudogenes of this gene have been identified throughout the human genome. [provided by RefSeq, Aug 2017] ACTB (Actin Beta) is a Protein Coding gene. Diseases associated with ACTB include Dystonia, Juvenile-Onset and Baraitser-Winter Syndrome 1 . Among its related pathways are Metabolism of proteins and Oxytocin signaling pathway . ACTB was the most highly expressed housekeeping gene. Keywords astrocytoma, glioblastoma multiforme, glioma, housekeeping gene, qPCR, quantitative real-time polymerase chain reaction, quantitative RT-PCR, reference gene, WHO grade Abbreviations Thus, it is essential to use reference genes that have been verified to be stably expressed within the specific experimental setting. used HKG’s such as Actb, B2M, and Gapdh. Perform up to 1000 assays (based on 20 µl assay volume) for each housekeeping gene. Housekeeping genes are typically constitutive genes that are required for the maintenance of basal cellular functions that are essential for the existence of a cell, regardless of its specific role in the tissue or organism. Biol. More specifically, ribosomal protein L27 performed favorably in human epicardial adipose tissue gene expression studies 14; similarly, ribosomal protein large PO (RPLPO) was shown to be the best housekeeping gene for expression studies during the differentiation of intestinal epithelial cells 15 or human myoblasts. However, one must still validate the chosen endogenous control or … β-Actin, a cytoskeletal protein, and GAPDH, an enzyme of glycolysis, are the two most widely used housekeeping genes, 1 on the basis of rather anecdotal evidence of … The housekeeping gene expression levels are fine-tuned to meet the metabolic requirements in various tissues. For experimental purposes, the expression of one or multiple housekeeping genes is used as a reference point for the analysis of expression levels of other genes. Comparison of the C T value of a target gene with that of the endogenous control gene allows the gene expression level of the target gene to be normalized to the amount of input RNA or cDNA. This is considered as one of the reproductive actins. Recent studies provide clear evidence that traditional non-validated "housekeeping" genes such as GAPDH and ACTB are not stably expressed in many experimental setups [27–30], and are therefore not suitable for normalization in a broad range of cell models. ACTB, GAPDH, and RPL13A showed different expression levels between astrozytoma grade II and primary glioblastoma. when HMBS and B2M were used as housekeeping genes for ACTB and 18S rRNA expression analysis in 3T3-L1 cells under normoxia (Average SD: 2.65 and … Beta-actin (ACTB) has traditionally been regarded as an endogenous housekeeping gene and has been widely used as a reference gene/protein in quantifying expression levels in tumors. UQCR2 was the most highly expressed following PSMB6 .
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