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Purified anti-TFEB Antibody

Pricing & Availability
Clone
W18197C (See other available formats)
Regulatory Status
RUO
Other Names
Transcription Factor EB
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
W18197C_PURE_TFEB_Antibody_1_021820
Whole cell extracts (15 µg protein) from NTERA-2 and HeLa (reduced expression controls), Ramos, Raji, Daudi and A20 cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:500 dilution) of purified anti-TFEB antibody (clone W18197C) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:50,000 dilution (lower). Lane M: Molecular weight marker. Cell lines were loaded in order of increasing predicted TFEB expression (source: Human Protein Atlas).
  • W18197C_PURE_TFEB_Antibody_1_021820
    Whole cell extracts (15 µg protein) from NTERA-2 and HeLa (reduced expression controls), Ramos, Raji, Daudi and A20 cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:500 dilution) of purified anti-TFEB antibody (clone W18197C) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:50,000 dilution (lower). Lane M: Molecular weight marker. Cell lines were loaded in order of increasing predicted TFEB expression (source: Human Protein Atlas).
  • W18197C_PURE_TFEB_Antibody_2_021820
    Whole cell extracts (250 µg total protein) prepared from Raji cells were immunoprecipitated overnight with 2.5 µg of purified rat IgG2a, κ isotype ctrl antibody (Cat. No. 400502) or purified anti-TFEB antibody (clone W18197C). The resulting IP fractions and whole cell extract input (6%) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with a control anti-TFEB antibody raised with a different immunogen. Lane M: Molecular weight marker.
  • W18197C_PURE_TFEB_Antibody_3_100721
    Daudi cells (filled histogram, positive control) and Jurkat cells (open histogram, negative control) were fixed and permeabilized using the True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401), and intracellularly stained with 0.125 µg/test of purified anti-TFEB antibody (clone W18197C) followed by PE goat anti-rat IgG antibody (Cat. No. 405406).
  • W18197C_PURE_TFEB_Antibody_4_121622
    IHC staining of Purified Anti-TFEB (clone W18197C) on formalin-fixed paraffin-embedded human colon tissue. Following antigen retrieval using Tris-EDTA (10mM Tris, 1mM EDTA, 0.05% Tween-20, pH 9.0), the tissue was incubated without (panel A) and with (panel B) 5 µg/mL of primary antibody followed by incubation with Alexa Fluor® 647 goat anti-rat IgG antibody (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 40X objective. Scale bar: 50 µm
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936101 25 µg £70
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936102 100 µg £174
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Description

The autophagy-lysosomal pathway is required for the quality control degradation of proteins and other macromolecules. The transcription factor EB (TFEB) plays a critical role in inducing expression of genes involved in autophagy and lysosomal biogenesis. The subcellular localization and activity of TFEB are regulated by mechanistic target of rapamycin (mTOR)-mediated phosphorylation. Under nutrient-replete conditions, mTORC1 phosphorylates TFEB promoting its cytosolic retention by 14-3-3 proteins and nuclear export through CRM1. Under nutrient deprivation, TFEB phosphorylation decreases, promoting its nuclear import and transactivation function. TFEB is frequently dysregulated in cancer, and is also involved in the clearance of intracellular pathogenic targets in a variety of diseases, such as Parkinson's and Alzheimer's, suggesting that novel therapeutic strategies could be based on the modulation of TFEB activity.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Partial recombinant human TFEB protein
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
IP, ICFC, IHC-P - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 1.0 µg/mL. For immunoprecipitation, the suggested use of this reagent is 2.5 µg/test. For intracellular flow cytometric staining, the suggested use of this reagent is ≤ 0.125 µg per million cells in 100 µL volume. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 5 - 10 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

When TFEB is phosphorylated, a molecular weight increase may be observed by western blot (Martina JA, et al. 2013. J Cell Biol. 200:475).

Differences in the observed molecular weight across cell lines and tissues may be attributed to differences in basal TFEB phosphorylation in these samples.

RRID
AB_2832906 (BioLegend Cat. No. 936101)
AB_2832906 (BioLegend Cat. No. 936102)

Antigen Details

Structure
TFEB is a 476 amino acid protein with a predicted molecular weight of 53 kD.
Distribution

Nucleo-cytoplasmic/Ubiquitously expressed

Function
Transcription Factor required for inducing expression of genes involved in autophagy and lysosomal biogenesis.
Biology Area
Cell Biology, Transcription Factors
Antigen References
  1. Napolitano G, et al. 2018. Nat Commun. 9:3312.
  2. Peña-Llopis S, et al. 2011. EMBO J. 30:3242.
  3. David R, et al. 2011. Nat Rev Mol Cell Biol. 12:404.
  4. Settembre C, et al. 2011. Science. 332:1429.
Gene ID
7942 View all products for this Gene ID
UniProt
View information about TFEB on UniProt.org

Related FAQs

There are no FAQs for this product.

Other Formats

View All TFEB Reagents Request Custom Conjugation
Description Clone Applications
Purified anti-TFEB W18197C WB,IP,ICFC,IHC-P
PE anti-TFEB W18197C ICFC
Alexa Fluor® 647 anti-TFEB W18197C ICFC
Go To Top Version: 3    Revision Date: 12/16/2022

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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