Recombinant Rabbit Anti-TFEB Antibody (clone R05-0D7) (CAT#: VS3-FY1439)

• Immunogen
• Human TFEB recombinant protein.

• Host Species
• Rabbit

• Type
• Rabbit IgG

• Specificity
• Human TFEB

• Species Reactivity
• Human

• Clone
• R05-0D7

• Applications
• Western Blot, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunocytochemistry, Immunofluorescence, Immunoprecipitation

• Conjugate
• Unconjugated

• MW
• Calculated MW: 53 kDa; Observed MW: 65-70 kDa

• Purification
• Affinity Purified

• Purity
• >95% as determined by SDS-PAGE

• Buffer
• 50 mM Tris-Glycine, pH 7.4, 0.15 M NaCl, 40% glycerol, 0.05% BSA

• Preservative
• 0.01% Sodium azide

• Storage
• Centrifuge briefly prior to opening vial. Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze/thaw cycles.

• Application Notes
• Western Blot: 1/500-1/1000
  Immunohistochemistry-Paraffin: 1/50-1/100
  Immunofluorescence: 1/50-1/200
  Immunoprecipitation: 1/20

• Alternative Names
• Transcription Factor EB; Class E Basic Helix-Loop-Helix Protein 35; BHLHE35; T-Cell Transcription Factor EB; ALPHATFEB; TCFEB

• Gene ID
• 7942

• UniProt ID
• P19484

• Sequence Similarities
• Belongs to the MiT/TFE family.

• Cellular Localization
• Cytoplasm, Lysosome, Membrane, Nucleus

• Post Translation Modifications
• Phosphorylation at Ser-211 by MTOR regulates its subcellular location and activity (PubMed:21617040, PubMed:22576015, PubMed:22343943, PubMed:22692423, PubMed:24081491, PubMed:30120233, PubMed:35662396).
  When nutrients are present, phosphorylation by MTOR promotes association with 14-3-3/YWHA adapters and retention in the cytosol (PubMed:22576015, PubMed:22343943, PubMed:22692423, PubMed:35662396).
  Inhibition of mTORC1, starvation and lysosomal disruption, promotes dephosphorylation and translocation to the nucleus (PubMed:22576015, PubMed:22343943, PubMed:22692423).
  Exported from the nucleus in a mTORC1-dependent manner in response to nutrient availability (PubMed:30120233).
  Dephosphorylated by phosphatase PPP3CA following Coxsackievirus B3 infection, leading to nuclear translocation (PubMed:33691586).
  Alkylated via a non-enzymatic covalent modification (PubMed:35662396).
  Itaconate, an anti-inflammatory metabolite generated in response to lipopolysaccharide, alkylates Cys-212, preventing association with 14-3-3/YWHA adapters, thereby promoting nuclear translocation and activity (PubMed:35662396).
  Sumoylated; does not affect dimerization with MITF.
  (Microbial infection) Cleavage by Coxsackievirus B3 protease 3C after site Gln-60. This non-phosphorylated cleavage product retains its ability to interact with TFEB, TFE3 or MITF and presents impaired transcriptional activity, resulting in disruption of lysosomal functions and increased viral infection.

• Protein Refseq
• NP_001161299.2; NP_001258872.1; NP_001258873.1

• Function
• Transcription factor that acts as a master regulator of lysosomal biogenesis, autophagy, lysosomal exocytosis, lipid catabolism, energy metabolism and immune response (PubMed:21617040, PubMed:22576015, PubMed:22343943, PubMed:22692423, PubMed:30120233, PubMed:31672913, PubMed:35662396).
  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 (PubMed:1748288, PubMed:19556463, PubMed:29146937).
  Involved in the cellular response to amino acid availability by acting downstream of MTOR: in the presence of nutrients, TFEB phosphorylation by MTOR promotes its cytosolic retention and subsequent inactivation (PubMed:21617040, PubMed:22576015, PubMed:22343943, PubMed:22692423, PubMed:35662396).
  Upon starvation or lysosomal stress, inhibition of MTOR induces TFEB dephosphorylation, resulting in nuclear localization and transcription factor activity (PubMed:22576015, PubMed:22343943, PubMed:22692423, PubMed:35662396).
  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, thereby playing a central role in expression of lysosomal genes (PubMed:19556463, PubMed:22692423).
  Regulates lysosomal positioning in response to nutrient deprivation by promoting the expression of PIP4P1 (PubMed:29146937).
  Acts as a positive regulator of autophagy by promoting expression of genes involved in autophagy (PubMed:21617040, PubMed:22576015, PubMed:23434374, PubMed:27278822).
  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 (By similarity).
  Specifically recognizes the gamma-E3 box, a subset of E-boxes, present in the heavy-chain immunoglobulin enhancer (PubMed:2115126).
  Plays a role in the signal transduction processes required for normal vascularization of the placenta (By similarity).
  Involved in the immune response to infection by the bacteria S.aureus, S.typhimurium or S.enterica: infection promotes itaconate production, leading to alkylation, resulting in nuclear localization and transcription factor activity (PubMed:35662396).
  Itaconate-mediated alkylation activates TFEB-dependent lysosomal biogenesis, facilitating the bacteria clearance during the antibacterial innate immune response (PubMed:35662396).

Secondary Antibody

• 2ND-054CQ
• Goat Anti-Rabbit IgG Antibody, R-PE

• 2ND-055CQ
• Chicken Anti-Rabbit IgG (H&L) Antibody

• 2ND-057CQ
• Donkey Anti-Rabbit IgG (H&L) Antibody

• 2ND-059CQ
• Goat Anti-Rabbit IgG (H&L) Antibody

• 2ND-060CQ
• Llama Anti-Rabbit IgG (H&L) Antibody

• 2ND-061CQ
• Sheep Anti-Rabbit IgG (H&L) Antibody

Isotype Control

• MOB-208CQ
• Rabbit IgG Isotype Control Antibody