TFEB Antibody | Affinity Biosciences

WB
Cites

1/7

Western blot analysis of TFEB using various lysates Lanes 1 - 2: Merged signal (red and green).

FIGURE 6 | Effect of RSV on TFEB activity in NRK-52E cells.

Fig.

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

FIGURE 5 Gastrodin increases the phosphorylation and nuclear translocation of FoxO1 to up‐regulate TFEB expression in the foam cells.

FIGURE 6 AMPK is a critical upstream regulator of FoxO1 and TFEB.

FIGURE 5 Expression and co-localization of Bax and Bcl2 in the rat dorsal CA1 region of hippocampus.

Product:	TFEB Antibody
Catalog:	AF7015
Description:	Rabbit polyclonal antibody to TFEB
Application:	WB IHC
Reactivity:	Human, Mouse, Rat
Prediction:	Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.:	55~70kDa; 53kD(Calculated).
Uniprot:	P19484
RRID:	AB_2835320

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

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Related Downloads

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

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.

Reactivity:

Human,Mouse,Rat

Prediction:

Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(80%), Xenopus(82%)

Clonality:

Polyclonal

Specificity:

TFEB Antibody detects endogenous levels of total TFEB.

RRID:

AB_2835320
Cite Format: Affinity Biosciences Cat# AF7015, RRID:AB_2835320.

Conjugate:

Unconjugated.

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Storage:

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.

Alias:

Fold/Unfold

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

Immunogens

Immunogen:

Uniprot:

P19484(TFEB_HUMAN) >>Visit HPA database.

Gene(ID):

TFEB(7942)

Description:

TFEB Probable transcription factor binds to the USF/MLTF site and probably recognizes E-box sequences in the heavy-chain immunoglobulin enhancer.

Sequence:

P19484 TFEB_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MASRIGLRMQLMREQAQQEEQRERMQQQAVMHYMQQQQQQQQQQLGGPPTPAINTPVHFQSPPPVPGEVLKVQSYLENPTSYHLQQSQHQKVREYLSETYGNKFAAHISPAQGSPKPPPAASPGVRAGHVLSSSAGNSAPNSPMAMLHIGSNPERELDDVIDNIMRLDDVLGYINPEMQMPNTLPLSSSHLNVYSSDPQVTASLVGVTSSSCPADLTQKRELTDAESRALAKERQKKDNHNLIERRRRFNINDRIKELGMLIPKANDLDVRWNKGTILKASVDYIRRMQKDLQKSRELENHSRRLEMTNKQLWLRIQELEMQARVHGLPTTSPSGMNMAELAQQVVKQELPSEEGPGEALMLGAEVPDPEPLPALPPQAPLPLPTQPPSPFHHLDFSHSLSFGGREDEGPPGYPEPLAPGHGSPFPSLSKKDLDLMLLDDSLLPLASDPLLSTMSPEASKASSRRSSFSMEEGDVL

Predictions

Predictions:

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

Chicken

Zebrafish

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P19484 As Substrate

P19484

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

Research Backgrounds

P19484_TFEB_HUMAN

Function:

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.

Subcellular Location:

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).

Subunit Structure:

Homodimer and heterodimer; with TFE3 or MITF.

Family&Domains:

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.

References

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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TFEB Antibody - #AF7015