WB
at-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.)).| Product Name | Phospho-TFEB (Ser142) Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | A synthesized peptide derived from human TFEB around the phosphorylation site of Ser142. |
| Modification | p-Ser142 |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | WB |
| WB:1:500-1:2000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | TFEB |
|---|---|
| Gene Synonyms | TCFEB BHLHE35 ALPHATFEB |
| Gene Full Name | transcription factor EB |
| Gene Summary | Enables DNA-binding transcription factor activity; enzyme binding activity; and transcription cis-regulatory region binding activity. Involved in several processes, including cellular response to amino acid starvation; lysosome localization; and positive regulation of autophagy. Located in cytosol; lysosomal membrane; and nucleoplasm. [provided by Alliance of Genome Resources, Apr 2022] |
| Molecular Weight(MW) | 55~70kDa; 53kD(Calculated). |
| Cellular Localization | Cytoplasm. Nucleus. |
WB
at-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.)).| Application Notes | WB:1:500-1:2000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at -20 °C. Stable for 12 months from date of receipt. |
| Storage Buffer | Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. |
