Product: Phospho-TFEB (Ser142) Antibody
Catalog: AF3845
Description: Rabbit polyclonal antibody to Phospho-TFEB (Ser142)
Application: WB
Reactivity: Human, Mouse, Rat
Mol.Wt.: 55~70kDa; 53kD(Calculated).
Uniprot: P19484
RRID: AB_2847159

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000
*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
Clonality:
Polyclonal
Specificity:
Phospho-TFEB (Ser142) Antibody detects endogenous levels of TFEB only when phosphorylated at Ser142.
RRID:
AB_2847159
Cite Format: Affinity Biosciences Cat# AF3845, RRID:AB_2847159.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
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:

A synthesized peptide derived from human TFEB around the phosphorylation site of Ser142.

Uniprot:
Gene(ID):
Sequence:
MASRIGLRMQLMREQAQQEEQRERMQQQAVMHYMQQQQQQQQQQLGGPPTPAINTPVHFQSPPPVPGEVLKVQSYLENPTSYHLQQSQHQKVREYLSETYGNKFAAHISPAQGSPKPPPAASPGVRAGHVLSSSAGNSAPNSPMAMLHIGSNPERELDDVIDNIMRLDDVLGYINPEMQMPNTLPLSSSHLNVYSSDPQVTASLVGVTSSSCPADLTQKRELTDAESRALAKERQKKDNHNLIERRRRFNINDRIKELGMLIPKANDLDVRWNKGTILKASVDYIRRMQKDLQKSRELENHSRRLEMTNKQLWLRIQELEMQARVHGLPTTSPSGMNMAELAQQVVKQELPSEEGPGEALMLGAEVPDPEPLPALPPQAPLPLPTQPPSPFHHLDFSHSLSFGGREDEGPPGYPEPLAPGHGSPFPSLSKKDLDLMLLDDSLLPLASDPLLSTMSPEASKASSRRSSFSMEEGDVL

PTMs - P19484 As Substrate

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)

Research Backgrounds

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).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
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). FGF21 ameliorates septic liver injury by restraining proinflammatory macrophages activation through the autophagy/HIF-1α axis. Journal of advanced research, 2024 (PubMed: 38599281) [IF=10.7]

Application: WB    Species: Mouse    Sample:

Fig. 3. LPS impaired autophagic flux in macrophages. (A, B) Representative immunofluorescence and quantification of iNOS-F4/80 double positive macrophages in liver from mice in indicated groups (n = 5). DAPI (blue), iNOS (red), and F4/80 (green). White arrows indicate iNOS-F4/80 double positive macrophages. Scale bars: 50 μm. (C, D) Representative immunofluorescence and quantification of LC3 puncta in F4/80-positive macrophages in liver from mice in indicated group. DAPI (blue), LC3β (red), and F4/80 (green) (n = 5). Scale bars: 50 μm. (E, F) Representative immunofluorescence and quantification of p62-F4/80 double positive macrophages in liver from mice in indicated groups. DAPI (blue), p62 (red), and F4/80 (green) (n = 5). White triangles indicate p62-F4/80 double positive macrophages. Scale bars: 50 μm. (G) The expression of LC3 and p62 in lysates of BMDMs in “control-group” and “LPS-group”, and normalized to β-actin (n = 4). (H) The expression of LC3 and p62 in lysates of BMDMs in indicated groups, and normalized to β-actin (n = 4). And Bafilomycin A1 was added for last 6 h. (I, J) Representative confocal images and quantification of Raw264.7 transduced with Lenti-GFP-mCherry-LC3B (n = 10). Scale bars: 20 μm. (K) The expression of LC3 and p62 in lysates of BMDMs in indicated groups, and normalized to β-actin (n = 4). 3-MA was given as pretreatment for 12 h before LPS administration. (L, M) Representative confocal images and quantification of Raw264.7 transduced with Lenti-GFP-LC3B in indicated groups (n = 8). Scale bars: 10 μm. (N) The expression of LC3 and p62 in lysates of BMDMs in indicated groups, and normalized to β-actin (n = 4). U0126, SP600125 and SB203580 were pretreated to BMDMs for 12 h. (O, P) Representative confocal images and quantification of Raw264.7 transduced with Lenti-GFP-LC3B in indicated groups (n = 8). Scale bars: 20 μm. (Q, R) Representative confocal images and quantification of pHLys Red in indicated groups (n = 5). Scale bars: 50 μm. (J) Real-time PCR analysis for lamp1, ctsa, ctsb, and ctsf from BMDMs in indicated group. All values are presented as mean ± SEM. Statistical significance was measured using the unpaired 2-tailed Student t test for two experimental groups and one way ANOVA test for multiple groups. ns = no significant. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

2). Tfeb-Mediated Transcriptional Regulation of Autophagy Induces Autosis during Ischemia/Reperfusion in the Heart. Cells, 2022 (PubMed: 35053374) [IF=6.0]

Application: WB    Species: rat    Sample: NRCMs

Tat-Beclin1-induced autosis is regulated by Tfeb modulation in NRCMs. (A–C) NRCMs were treated with 5 µM Scrambled or Tat-Beclin 1 for 6 h and analyzed via Western blotting using anti-Tfeb, anti-pTfeb (Ser142) and anti-Gapdh antibodies (A) The relative ratios of Tfeb to Gapdh (B) and pTfeb to Tfeb (C) were quantified (mean values ± S.E., n = 6 for Tfeb, n = 3 for p-Tfeb; * p < 0.05, *** p < 0.001). (D,E) NRCMs were transduced with either Ad-lacZ or Ad-Tfeb for 24 h (D) or Ad-lacZ or Ad-shTfeb for 48 h (E) and then treated with 5 µM Scrambled or 0.5, 1 or 5 µM Tat-Beclin 1 for 3 h. Cell death was quantified via CellTiter-Blue assays (mean values ± S.D., n = 16; * p < 0.05, ** p < 0.01, not significant (n.s.)) (D,E). (F,G) Knockdown of Tfeb by Ad-shTfeb was confirmed via immunoblot analyses, using anti-Tfeb and anti-Gapdh antibodies (F). Knockdown of Atg7 by Ad-shAtg7 was confirmed via immunoblot analyses, using anti-Atg7 and anti-Gapdh antibodies (G). (H) NRCMs were transduced with either Ad-lacZ or Ad-Tfeb with or without Ad-shAtg7 as indicated for 48 h and then treated with 5 µM Scrambled or Tat-Beclin 1 for 3 h. Cell death was quantified via CellTiter-Blue assays (mean values ± S.D., n = 16; * p < 0.05, ** p < 0.01, not significant (n.s.)).

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