Phospho-TFEB (Ser211) Antibody | Affinity Biosciences

IHC
IF/ICC
Cites

1/6

AF3708 at 1/100 staining Human kidney cancer and adjacent normal tissues by IHC-P.

AF3708 at 1/100 staining Human kidney cancer by IHC-P.

AF3708 staining A549 cells(4h of LPS treatment) by IF/ICC.

Fig.

FIGURE 6 TS treatment promoted autophagy and clear p-tau.

Product:	Phospho-TFEB (Ser211) Antibody
Catalog:	AF3708
Description:	Rabbit polyclonal antibody to Phospho-TFEB (Ser211)
Application:	WB IHC IF/ICC
Reactivity:	Human, Mouse, Rat
Mol.Wt.:	55~70kDa; 53kD(Calculated).
Uniprot:	P19484
RRID:	AB_2847022

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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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IHC Protocol

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Product Info

Source:

Rabbit

Application:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*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 (Ser211) Antibody detects endogenous levels of TFEB only when phosphorylated at Ser211.

RRID:

AB_2847022
Cite Format: Affinity Biosciences Cat# AF3708, RRID:AB_2847022.

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 Ser211.

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). Cyclic helix B peptide alleviates proinflammatory cell death and improves functional recovery after traumatic spinal cord injury. Redox Biology (PubMed: 37290302) [IF=11.4]

2). FGF21 augments autophagy in random-pattern skin flaps via AMPK signaling pathways and improves tissue survival. Cell Death & Disease (PubMed: 31740658) [IF=9.0]

Application: WB Species: Rat Sample: Fap tissue

Fig. 7 FGF21 boosted autophagy via enhancing TFEB activity. On the 7th day after operation, samples were harvested from the Control and FGF21 groups to evaluate the TFEB level. Further, transfection of the AVV TFEB shRNA was performed to inhibit the expression of TFEB in the cells, and then the expression of relative proteins was compared in the FGF21, Scramble and TFEB shRNA group. a Immunofluorescence exhibiting more expression of TFEB in the FGF21 group than that in the Control group (scan bar, 15 μm). b Histogram showing the percentage of TFEB translocation into nucleus in dermal layer. c Western blotting showing levels of P-TFEB (Ser221) and nuclear TFEB. d Histogram showing quantificational comparison of cytoplasmic and nuclear TFEB expressions between the Control and FGF21 groups. e Immunofluorescence exhibiting the expression of LC3II in the FGF21, Scramble and TFEB shRNA groups. f Quantification of percentage of positive cells with LC3II labeled autophagosomes in dermal layer. g Western blotting showing levels of P-TFEB (Ser221) and nuclear TFEB in the FGF21, Scramble and TFEB shRNA groups. h Quantification of cytoplasmic and nuclear TFEB expressions detected by western blotting. i Western blotting showing levels of autophagic proteins (p62, Beclin1, VPS34, CTSD, and LC3II) in the FGF21, Scramble and TFEB shRNA groups, which was corrected by GAPDH as internal control. j Quantification of expression of autophagy-related proteins (p62, Beclin1, VPS34, CTSD, and LC3II) in western blotting. Significance: *p < 0.05 and **p < 0.01 vs the Control group. #p < 0.05 and ##p < 0. 01 vs the Scramble group. Data were expressed as means ± SEM (n = 6 per group).

3). mTOR deletion ameliorates CD4+ T cell apoptosis during sepsis by improving autophagosome-lysosome fusion. APOPTOSIS (PubMed: 35435531) [IF=7.2]

Application: WB Species: mouse Sample: CD4+T cells

Fig. 3| Transcription factor EB (TFEB) phosphorylation and SNARE expression in septic mice. (A, a–e) Protein levels of phospho (p)-TFEB, syntaxin 17 (STX17), vesicle associated membrane protein 8 (VAMP8), and synaptosomal-associated protein 29 (SNAP29) in CD4+T cells were quantifed by western blotting in wild-type (WT)sham-treated mice, WT CLP mice, mammalian target of rapamycin(mTOR) knockout via Cre expression under the control of lymphocyte-specifc protein tyrosine kinase (Lck) (Lck-mTOR) CLP mice,and tuberous sclerosis complex 1 (TSC1) knockout (Lck-TSC1) CLP mice.

4). Therapeutic effects of total saikosaponins from Radix bupleuri against Alzheimer’s disease. Frontiers in Pharmacology (PubMed: 35935875) [IF=5.6]

Application: WB Species: Mice Sample: PC12 cells

FIGURE 6 TS treatment promoted autophagy and clear p-tau. (A) The effect of TS on p-tau (Ser 396), NDP52, p62 and LC3-II and Beclin-1 proteins in the Aβ-induced PC12 cells were detected by Western blot. (B) Quantitative analysis of the relative expression concertation of p-tau (Ser 396) protein in each group (# p = 0.0409, *p = 0.0130, **p = 0.0029). (C) Quantitative analysis of the relative expression concertation of NDP52 protein in each group (# p = 0.0113, **p = 0.0057). (D) Quantitative analysis of the gray value of p62 protein bands in each group (## p = 0.0093, *p = 0.0312, **p = 0.0084, **p = 0.0093, **p = 0.0047). (E) Quantitative analysis of the relative expression concertation of LC3-II protein in each group (#### p < 0.0001, **p = 0.0053, **p = 0.0037). (F) Quantitative analysis of the relative expression concertation of Beclin-1 protein in each group (### p = 0.0002, *p = 0.0356). (G) Quantitative analysis of the relative expression level of mTOR protein in each group (# p = 0.0450, *p = 0.0359, **p = 0.0068). (H) Quantitative analysis of the relative expression level of p-TFEB protein in each group (# p = 0.0435, *p = 0.0129, **p = 0.0099). #Compared with WT group; *Compared with model group. Data are presented as mean ± SEM (n = 3).

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Phospho-TFEB (Ser211) Antibody - #AF3708