Phospho-TFEB (Ser142) Antibody | Affinity Biosciences

Cites

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Tat-Beclin1-induced autosis is regulated by Tfeb modulation in NRCMs.

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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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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MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

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:

P19484(TFEB_HUMAN) >>Visit HPA database.

Gene(ID):

TFEB(7942)

Sequence:

P19484 TFEB_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MASRIGLRMQLMREQAQQEEQRERMQQQAVMHYMQQQQQQQQQQLGGPPTPAINTPVHFQSPPPVPGEVLKVQSYLENPTSYHLQQSQHQKVREYLSETYGNKFAAHISPAQGSPKPPPAASPGVRAGHVLSSSAGNSAPNSPMAMLHIGSNPERELDDVIDNIMRLDDVLGYINPEMQMPNTLPLSSSHLNVYSSDPQVTASLVGVTSSSCPADLTQKRELTDAESRALAKERQKKDNHNLIERRRRFNINDRIKELGMLIPKANDLDVRWNKGTILKASVDYIRRMQKDLQKSRELENHSRRLEMTNKQLWLRIQELEMQARVHGLPTTSPSGMNMAELAQQVVKQELPSEEGPGEALMLGAEVPDPEPLPALPPQAPLPLPTQPPSPFHHLDFSHSLSFGGREDEGPPGYPEPLAPGHGSPFPSLSKKDLDLMLLDDSLLPLASDPLLSTMSPEASKASSRRSSFSMEEGDVL

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). Tfeb-Mediated Transcriptional Regulation of Autophagy Induces Autosis during Ischemia/Reperfusion in the Heart. Cells (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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Phospho-TFEB (Ser142) Antibody - #AF3845