TFEB Antibody - #AF7015

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製品: TFEB Antibody
カタログ: AF7015
タンパク質の説明: Rabbit polyclonal antibody to TFEB
アプリケーション: WB IHC
反応性: Human, Mouse, Rat
予測: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
分子量: 55~70kDa; 53kD(Calculated).
ユニプロット: P19484
RRID: AB_2835320

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MSDS
説明書
COA
プロトコル
WB Handbook
IHC Protocol
IF/ICC Protocol

製品説明

ソース:
Rabbit
アプリケーション:
WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

反応性:
Human,Mouse,Rat
予測:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(80%), Xenopus(82%)
クローナリティ:
Polyclonal
特異性:
TFEB Antibody detects endogenous levels of total TFEB.
RRID:
AB_2835320
引用形式: Affinity Biosciences Cat# AF7015, RRID:AB_2835320.
コンジュゲート:
Unconjugated.
精製:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
保存:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
別名:

折りたたみ／展開

Alpha TFEB; AlphaTFEB; bHLHe35; Class E basic helix-loop-helix protein 35; T cell transcription factor EB; TCFEB; TFEB; TFEB_HUMAN; Transcription factor EB;

免疫原

免疫原:
Uniprot:

P19484(TFEB_HUMAN) >>Visit HPA database.

遺伝子(ID):
TFEB(7942)
タンパク質の説明:
TFEB Probable transcription factor binds to the USF/MLTF site and probably recognizes E-box sequences in the heavy-chain immunoglobulin enhancer.
タンパク質配列:
MASRIGLRMQLMREQAQQEEQRERMQQQAVMHYMQQQQQQQQQQLGGPPTPAINTPVHFQSPPPVPGEVLKVQSYLENPTSYHLQQSQHQKVREYLSETYGNKFAAHISPAQGSPKPPPAASPGVRAGHVLSSSAGNSAPNSPMAMLHIGSNPERELDDVIDNIMRLDDVLGYINPEMQMPNTLPLSSSHLNVYSSDPQVTASLVGVTSSSCPADLTQKRELTDAESRALAKERQKKDNHNLIERRRRFNINDRIKELGMLIPKANDLDVRWNKGTILKASVDYIRRMQKDLQKSRELENHSRRLEMTNKQLWLRIQELEMQARVHGLPTTSPSGMNMAELAQQVVKQELPSEEGPGEALMLGAEVPDPEPLPALPPQAPLPLPTQPPSPFHHLDFSHSLSFGGREDEGPPGYPEPLAPGHGSPFPSLSKKDLDLMLLDDSLLPLASDPLLSTMSPEASKASSRRSSFSMEEGDVL

種類予測

種類予測:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Rabbit
100
Xenopus
82
Chicken
80
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P19484 基板として

Site PTM Type Enzyme
S3 Phosphorylation
R8 Methylation
S74 Phosphorylation
K91 Ubiquitination
Y95 Phosphorylation
S97 Phosphorylation
Y100 Phosphorylation
S109 Phosphorylation
S114 Phosphorylation
K116 Acetylation
S122 Phosphorylation P42345 (MTOR)
S133 Phosphorylation
S134 Phosphorylation
S138 Phosphorylation
S142 Phosphorylation P28482 (MAPK1) , P42345 (MTOR)
T183 Phosphorylation
S211 Phosphorylation P42345 (MTOR)
S227 Phosphorylation
R254 Methylation
K274 Ubiquitination
T330 Phosphorylation
T331 Phosphorylation
S332 Phosphorylation
S334 Phosphorylation
S423 Phosphorylation
S429 Phosphorylation
S441 Phosphorylation
S455 Phosphorylation
S462 Phosphorylation P05771 (PRKCB)
S463 Phosphorylation P05771 (PRKCB)
S466 Phosphorylation
S467 Phosphorylation P05771 (PRKCB)
S469 Phosphorylation P05771 (PRKCB)

研究背景

機能:

Transcription factor that specifically recognizes and binds E-box sequences (5'-CANNTG-3'). Efficient DNA-binding requires dimerization with itself or with another MiT/TFE family member such as TFE3 or MITF. In association with TFE3, activates the expression of CD40L in T-cells, thereby playing a role in T-cell-dependent antibody responses in activated CD4(+) T-cells and thymus-dependent humoral immunity. Specifically recognizes and binds the CLEAR-box sequence (5'-GTCACGTGAC-3') present in the regulatory region of many lysosomal genes, leading to activate their expression. It thereby plays a central role in expression of lysosomal genes. Acts as a positive regulator of autophagy by promoting expression of genes involved in autophagy. Specifically recognizes the gamma-E3 box, a subset of E-boxes, present in the heavy-chain immunoglobulin enhancer. Plays a role in the signal transduction processes required for normal vascularization of the placenta. Regulates lysosomal positioning in response to nutrient deprivation by promoting the expression of PIP4P1.

PTMs:

Sumoylated; does not affect dimerization with MITF.

細胞の位置付け:

Cytoplasm. Nucleus.
Note: Mainly present in the cytoplasm (PubMed:23434374). Under aberrant lysosomal storage conditions, it translocates from the cytoplasm to the nucleus (PubMed:23434374). The translocation to the nucleus is regulated by ATP13A2 (PubMed:23434374, PubMed:27278822). In macrophages, translocates into the nucleus upon live S.enterica infection (PubMed:27184844).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
サブユニット構造:

Homodimer and heterodimer; with TFE3 or MITF.

タンパク質ファミリー:

The leucin zipper region is essential for homo- or heterodimerization and high-affinity DNA binding. DNA binding is mediated by the basic region.

Belongs to the MiT/TFE family.

参考文献

1). Morphine-induced microglial immunosuppression via activation of insufficient mitophagy regulated by NLRX1. Journal of Neuroinflammation, 2022 (PubMed: 35414088) [IF=9.3]
2). Resveratrol Attenuates Oxalate-Induced Renal Oxidative Injury and Calcium Oxalate Crystal Deposition by Regulating TFEB-Induced Autophagy Pathway. Frontiers in Cell and Developmental Biology, 2021 (PubMed: 33718378) [IF=5.5]

Application: IF/ICC    Species: rat    Sample: nrK-52E cell

FIGURE 6 | Effect of RSV on TFEB activity in NRK-52E cells.(C) TFEB immunofluorescence assay results showed that the fluorescence intensity of TFEB antibody in nrK-52E cell nucleus increased after RSV treatment. *P < 0.05, **P < 0.01.

Application: WB    Species: rat    Sample: nrK-52E cell

FIGURE 6 | Effect of RSV on TFEB activity in NRK-52E cells.(A) NRK-52E cells were treated with RSV at different concentrations (0–32 µM) for 24 h. The expressions of nuclear TFEB (n-TFEB), cytoplasmic TFEB (c-TFEB), and total TFEB (t-TFEB) proteins were detected by western blot assay. Quantitative analysis of the densities of western blot bands.
3). Gastrodin induces lysosomal biogenesis and autophagy to prevent the formation of foam cells via AMPK‐FoxO1‐TFEB signalling axis. Journal of Cellular and Molecular Medicine, 2021 (PubMed: 33973365) [IF=5.3]

Application: WB    Species: Mice    Sample: foam cells

FIGURE 5 Gastrodin increases the phosphorylation and nuclear translocation of FoxO1 to up‐regulate TFEB expression in the foam cells. A, Gastrodin increased the phosphorylation of FoxO1 in the foam cells. B and C, Gastrodin promoted nuclear translocation of FoxO1 in the foam cells. B, Images of the subcellular locations of FoxO1. C, Representative blots of cytoplasmic and nuclear fractions of FoxO1. D and E, The determination of mRNA and protein expression levels of FoxO1 after transfecting by control shRNA and shFoxO1. D, Measurement of mRNA expression level by RT‐PCR. E, Measurement of protein expression level by Western blotting. F and G, Inhibition of FoxO1 expression abolished the effect of gastrodin on autophagy in the foam cells. The protein levels of LC3I/II and p62 F, and mRNA levels of autophagic genes G, were determined by Western blotting and RT‐PCR. H, Inhibition of FoxO1 reduced the protein level of TFEB. *P < .05; **P < .01; ***P < .001. Results are presented as mean ±SD of three independent experiments. The value represents fold of vehicle. shCont., Control shRNA. Cyto, Cytoplasm; Nucl, Nucleus

Application: IF/ICC    Species: Mice    Sample: foam cells

FIGURE 6 AMPK is a critical upstream regulator of FoxO1 and TFEB. A and B, Gastrodin activated AMPK in the foam cells. A, Representative blots of AMPK and p‐AMPK in macrophages. B, Immunofluorescence analysis of p‐AMPK in macrophages. C and D, The inhibition of AMPK activity decreased the phosphorylation of FoxO1 and nuclear translocation of TFEB. Macrophages were treated with CC (10μM) for 1 h. The phosphorylation level of FoxO1 was analysed by Western blotting C, and nuclear translocation of TFEB was determined by immunofluorescence D. *P < .05; **P < .01. Results are presented as mean ± SD of three independent experiments. The value represents fold of vehicle. CC, Dorsomorphin dihydrochloride
4). Puerarin Alleviates Vascular Cognitive Impairment in Vascular Dementia Rats. Frontiers in Behavioral Neuroscience, 2021 (PubMed: 34720898) [IF=3.0]

Application: WB    Species: Rat    Sample: hippocampus

FIGURE 5 Expression and co-localization of Bax and Bcl2 in the rat dorsal CA1 region of hippocampus. (A) Protein expression of Bax in the hippocampal dorsal CA1 region of rats. (B) Protein expression of Bcl2 in the hippocampal dorsal CA1 region of rats. (C) Bax/Bcl2 ratios in the hippocampal dorsal CA1 region of rats. (D) Pur’s effects on Bax and Bcl2 co-localization in the hippocampal dorsal CA1 region of rats, assessed by immunofluorescent double staining. Representative micrographs are shown at ×200. Results were mean ± SEM (n = 6). *P < 0.05, **P < 0.01 vs. Sham; ##P < 0.01 vs. BCCAO; $P < 0.05, $$P < 0.01 vs. BCCAO + Pur (150 mg/kg/day); &P < 0.05, &&P < 0.01 vs. BCCAO + NGF (20 mg/kg/day).
5). Transcription factor EB promotes rheumatoid arthritis of Sprague–Dawley rats via regulating autophagy. 3 Biotech, 2021 (PubMed: 33786279) [IF=2.8]

Application: IHC    Species: Rat    Sample: RA rats

Fig. 2 Changes of autophagy-related genes in RA rats. The expressions of transcription factor EB (TFEB), microtubule-associated proteins 1 light chain 3 (LC3B), and p62 in the synovial tissues by IHC (n = 3, biological replications). Original magnification ×150

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