TATA box-binding protein-like protein 1 is a protein that in humans is encoded by the TBPL1 gene. Initiation of transcription by RNA polymerase II requires the activities of more than 70 polypeptides. The protein that coordinates these activities is transcription factor IID (TFIID), which binds to the core promoter to position the polymerase properly, serves as the scaffold for assembly of the remainder of the transcription complex, and acts as a channel for regulatory signals. TFIID is composed of the TATA-binding protein (TBP) and a group of evolutionarily conserved proteins known as TBP-associated factors or TAFs. TAFs may participate in basal transcription, serve as coactivators, function in promoter recognition or modify general transcription factors (GTFs) to facilitate complex assembly and transcription initiation. This gene encodes a protein that serves the same function as TBP and substitutes for TBP at some promoters that are not recognized by TFIID. It is essential for spermiogenesis and believed to be important in expression of developmentally regulated genes.
Background References
1. Qin H et al. C-MYC induces idiopathic pulmonary fibrosis via modulation of miR-9-5p-mediated TBPL1. Cell Signal. 2022 May
2. Mishal R et al. Role of the TATA-box binding protein (TBP) and associated family members in transcription regulation. Gene. 2022 Jul
Western blot analysis of TBPL1 on different lysates with Rabbit anti-TBPL1 antibody (HA722426) at 1/1,000 dilution.
Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: 293T cell lysate (20 µg/Lane) Lane 3: Mouse testis tissue lysate (40 µg/Lane) Lane 4: Rat testis tissue lysate (40 µg/Lane)
Predicted band size: 21 kDa Observed band size: 25 kDa
Exposure time: 20 second; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722426) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-TBPL1 antibody (HA722426) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722426) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-TBPL1 antibody (HA722426) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722426) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-TBPL1 antibody (HA722426) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722426) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Flow cytometric analysis of 293T cells labeling TBPL1.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA722426, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
TBPL1 was immunoprecipitated from 0.2 mg 293T cell lysate with HA722426 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722426 at 1/1,000 dilution. Mouse anti Rabbit IgG heavy chain (Fc) secondary antibody (M1003-7) at 1/50,000 dilution was used for 1 hour at room temperature.
Lane 1: 293T cell lysate (input) Lane 2: HA722426 IP in 293T cell lysate Lane 3: Rabbit IgG instead of HA722426 in 293T cell lysate