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Transcription Factor EB Antibody (YA5876)

(Synonyms: BHLHE35, TFEB, Transcription factor EB, Class E basic helix-loop-helix protein 35, bHLHe35)
Customer Review

Based on 1 Customer Validation

Transcription Factor EB Antibody (YA5876) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Transcription Factor EB.

For research use only. We do not sell to patients.
  • Host:

    Rabbit

  • Isotype:

    IgG

  • Application:

    WB, IHC-P, ICC/IF, IP, ELISA

  • Reactivity :

    Human, Mouse, Rat

  • Formulation:

    Supplied in PBS, 50% glycerol, 0.05% Proclin 300, 0.05%BSA

  • Conjugation:
    Non-conjugated
ALL WB ICC
  • Experimental Validation Results for Transcription Factor EB Antibody (YA5876)
  • Experimental Validation Results for Transcription Factor EB Antibody (YA5876)
  • Experimental Validation Results for Transcription Factor EB Antibody (YA5876)
  • Transcription Factor EB Antibody (YA5876) Product Image
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Tel: 609-228-6898 E-mail: [email protected] [email protected]

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Use of MCE Products

Applications

Application
IHC-P Info
IHC-P: Immunohistochemistry-Paraffin
WB Info
WB: Western Blot
ICC/IF Info
ICC/IF: Immunocytochemistry/
Immunofluorescence
ELISA Info
ELISA: Enzyme Linked Immunosorbent Assay
IP Info
IP: Immunoprecipitation
Dilution Ratio 1:200-1:1000 1:2000-1:10000 1:200-1:1000 1:5000-1:20000 1:50-1:200

Product Details

Description

Transcription Factor EB Antibody (YA5876) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Transcription Factor EB.

  • Host Rabbit
  • Clonality Monoclonal
  • Species Reactivity
    Human, Mouse, Rat
  • Observed Molecular Weight
    Observed band size: 65-70 kDa Info
    Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
  • Calculated Molecular Weight Predicted band size: 53 kDa
Purification

Protein A

Conjugation

Non-conjugated

Modification

Unmodified

Isotype

IgG

RRID

AB_3719100

Product Properties

  • Appearance

    Solution

  • Formulation

    Supplied in PBS, 50% glycerol, 0.05% Proclin 300, 0.05%BSA

  • Concentration

    Batch-dependent, Please check the COA for the concentration of each lot. Check Lot Concentration

  • Storage & Stability

    Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

  • Shipping

    Shipping with blue ice.

Verification Images

  • Experimental Validation Results for Transcription Factor EB Antibody (YA5876)
    Western blot analysis of extracts from Raji (lane 2(20μg), HeLa (lane 3(20μg), RAW264 (lane 4(20μg) and A549 (lane 5(20μg) using Transcription Factor EB Antibody (HY-P86184) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000)and Loading control antibody (Beta Actin, HY-P80993, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
  • Experimental Validation Results for Transcription Factor EB Antibody (YA5876)
    Immunocytochemistry analysis of Hela cells labeling Transcription Factor EB with Transcription Factor EB Antibody (HY-P86184) at 1/300 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with Transcription Factor EB Antibody (HY-P86184) at 1/300 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for Transcription Factor EB Antibody (YA5876)
    Immunocytochemistry analysis of Hela cells labeling Transcription Factor EB with Transcription Factor EB Antibody (HY-P86184) at 1/600 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with Transcription Factor EB Antibody (HY-P86184) at 1/600 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

Background

  • Function

    Transcription Factor EB transcription factor that acts as a master regulator of lysosomal biogenesis, autophagy, lysosomal exocytosis, lipid catabolism, energy metabolism and immune response. 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. 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. Upon starvation or lysosomal stress, inhibition of MTOR induces TFEB dephosphorylation, resulting in nuclear localization and transcription factor activity. 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. Regulates lysosomal positioning in response to nutrient deprivation by promoting the expression of PIP4P1. Acts as a positive regulator of autophagy by promoting expression of genes involved in autophagy. 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 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. 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. Itaconate-mediated alkylation activates TFEB-dependent lysosomal biogenesis, facilitating the bacteria clearance during the antibacterial innate immune response. In association with ACSS2, promotes the expression of genes involved in lysosome biogenesis and both autophagy upon glucose deprivation[1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20].

  • Subcellular Localization

    Nucleus; Cytoplasm, cytosol; Lysosome membrane; Nucleus

  • Isoforms & Post-Translational Modification

    P19484 has 2 isomers: P19484-1: 52865 Da (predicted); P19484-2: 43803 Da (predicted).
    Phosphorylation at Ser-211 by MTOR via non-canonical mTORC1 pathway regulates its subcellular location and activity (PubMed:21617040, PubMed:22343943, PubMed:22576015, PubMed:22692423, PubMed:23401004, PubMed:24081491, PubMed:25720963, PubMed:30120233, PubMed:32612235, PubMed:32753672, PubMed:35662396, PubMed:36697823). When nutrients are present, phosphorylation by MTOR promotes association with 14-3-3/YWHA adapters and retention in the cytosol (PubMed:22343943, PubMed:22576015, PubMed:22692423, PubMed:23401004, PubMed:25720963, PubMed:32612235, PubMed:32753672, PubMed:35662396). Inhibition of mTORC1, starvation and lysosomal disruption, promotes dephosphorylation by calcineurin PPP3CB and translocation to the nucleus (PubMed:22343943, PubMed:22576015, PubMed:22692423, PubMed:25720963, PubMed:32753672). Dephosphorylated by calcineurin PPP3CB in response to lysosomal Ca(2+) release (PubMed:25720963). IRGM promotes dephosphorylation by calcineurin PPP3CB, resulting in TFEB nuclear translocation and stimulation of lysosomal biogenesis (PubMed:32753672). Dephosphorylated by phosphatase PPP3CA following Coxsackievirus B3 infection, leading to nuclear translocation (PubMed:33691586). Exported from the nucleus in a mTORC1-dependent manner in response to nutrient availability (PubMed:30120233);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

  • Subunit

    Homodimer and heterodimer; with TFE3 or MITF (PubMed:15507434, PubMed:1748288). Interacts (when phosphorylated by MTOR) with YWHAZ; promoting retention in the cytosol (PubMed:35662396). Interacts with IRGM; promoting association between TFEB and PPP3CB and dephosphorylation (PubMed:32753672). Interacts with small GTPases Rag (RagA/RRAGA, RagB/RRAGB, RagC/RRAGC and/or RagD/RRAGD); promoting its recruitment to lysosomal membrane in the presence of nutrients (PubMed:23401004). Interacts with ACSS2 (PubMed:28552616)

  • SwissProt ID

    P19484

  • Gene ID
    7942 [NCBI]
  • Synonyms

    BHLHE35, TFEB, Transcription factor EB, Class E basic helix-loop-helix protein 35, bHLHe35

Transcription Factor EB Antibody (YA5876) Related Classifications

MOQ
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100 mg

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

    MedChemExpress Validation
     Western blot analysis of extracts from Raji (lane 2(20μg), HeLa (lane 3(20μg), RAW264 (lane 4(20μg) and A549 (lane 5(20μg) using Transcription Factor EB Antibody (HY-P86184) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000)and Loading control antibody (Beta Actin, HY-P80993, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.

  • 1/16

    MedChemExpress Validation
     Immunocytochemistry analysis of Hela cells labeling Transcription Factor EB with Transcription Factor EB Antibody (HY-P86184) at 1/300 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with Transcription Factor EB Antibody (HY-P86184) at 1/300 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • 1/16

    MedChemExpress Validation
     Immunocytochemistry analysis of Hela cells labeling Transcription Factor EB with Transcription Factor EB Antibody (HY-P86184) at 1/600 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with Transcription Factor EB Antibody (HY-P86184) at 1/600 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • Transcription Factor EB Antibody (YA5876) Product Image
    1/16
  • Experimental Validation Results for Transcription Factor EB Antibody (YA5876)
  • Experimental Validation Results for Transcription Factor EB Antibody (YA5876)
  • Experimental Validation Results for Transcription Factor EB Antibody (YA5876)
  • Transcription Factor EB Antibody (YA5876) Product Image
  • Experimental Validation Results for Transcription Factor EB Antibody (YA5876)
    1/1
    MedChemExpress Validation
     Western blot analysis of extracts from Raji (lane 2(20μg), HeLa (lane 3(20μg), RAW264 (lane 4(20μg) and A549 (lane 5(20μg) using Transcription Factor EB Antibody (HY-P86184) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000)and Loading control antibody (Beta Actin, HY-P80993, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
  • Experimental Validation Results for Transcription Factor EB Antibody (YA5876)
    1/1
    MedChemExpress Validation
     Immunocytochemistry analysis of Hela cells labeling Transcription Factor EB with Transcription Factor EB Antibody (HY-P86184) at 1/300 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with Transcription Factor EB Antibody (HY-P86184) at 1/300 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for Transcription Factor EB Antibody (YA5876)
    1/1
    MedChemExpress Validation
     Immunocytochemistry analysis of Hela cells labeling Transcription Factor EB with Transcription Factor EB Antibody (HY-P86184) at 1/600 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with Transcription Factor EB Antibody (HY-P86184) at 1/600 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
  • Experimental Validation Results for Transcription Factor EB Antibody (YA5876)
  • Experimental Validation Results for Transcription Factor EB Antibody (YA5876)
  • Experimental Validation Results for Transcription Factor EB Antibody (YA5876)
  • Transcription Factor EB Antibody (YA5876) Product Image
    MedChemExpress Validation

    Western blot analysis of extracts from Raji (lane 2(20μg), HeLa (lane 3(20μg), RAW264 (lane 4(20μg) and A549 (lane 5(20μg) using Transcription Factor EB Antibody (HY-P86184) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000)and Loading control antibody (Beta Actin, HY-P80993, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.

  • Transcription Factor EB Antibody (YA5876) Product Image
    MedChemExpress Validation

    Immunocytochemistry analysis of Hela cells labeling Transcription Factor EB with Transcription Factor EB Antibody (HY-P86184) at 1/300 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with Transcription Factor EB Antibody (HY-P86184) at 1/300 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • Transcription Factor EB Antibody (YA5876) Product Image
    MedChemExpress Validation

    Immunocytochemistry analysis of Hela cells labeling Transcription Factor EB with Transcription Factor EB Antibody (HY-P86184) at 1/600 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with Transcription Factor EB Antibody (HY-P86184) at 1/600 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • Transcription Factor EB Antibody (YA5876) Product Image WB
  • Transcription Factor EB Antibody (YA5876) Product Image ICC
  • Transcription Factor EB Antibody (YA5876) Product Image ICC