Syntaxin-4 is a protein that in humans is encoded by the STX4 gene. Plasma membrane t-SNARE that mediates docking of transport vesicles (By similarity). Necessary for the translocation of SLC2A4 from intracellular vesicles to the plasma membrane (By similarity). In neurons, recruited at neurite tips to membrane domains rich in the phospholipid 1-oleoyl-2-palmitoyl-PC (OPPC) which promotes neurite tip surface expression of the dopamine transporter SLC6A3/DAT by facilitating fusion of SLC6A3-containing transport vesicles with the plasma membrane (By similarity). Together with STXB3 and VAMP2, may also play a role in docking/fusion of intracellular GLUT4-containing vesicles with the cell surface in adipocytes and in docking of synaptic vesicles at presynaptic active zones (By similarity).
Background References
1. Black HL et al. Knockout of syntaxin-4 in 3T3-L1 adipocytes reveals new insight into GLUT4 trafficking and adiponectin secretion. J Cell Sci. 2022 Dec
2. Schrauwen I et al. Syntaxin 4 is essential for hearing in human and zebrafish. Hum Mol Genet. 2023 Mar
Western blot analysis of Syntaxin 4 on different lysates with Rabbit anti-Syntaxin 4 antibody (HA721509) at 1/1,000 dilution.
Lane 1: 293T cell lysate (20 µg/Lane) Lane 2: HeLa cell lysate (20 µg/Lane) Lane 3: A431 cell lysate (20 µg/Lane) Lane 4: A549 cell lysate (20 µg/Lane) Lane 5: K-562 cell lysate (20 µg/Lane) Lane 6: MCF7 cell lysate (20 µg/Lane) Lane 7: HCT 116 cell lysate (20 µg/Lane) Lane 8: U-87 MG cell lysate (20 µg/Lane) Lane 9: HUVEC cell lysate (20 µg/Lane) Lane 10: NIH/3T3 cell lysate (20 µg/Lane) Lane 11: RAW264.7 cell lysate (20 µg/Lane) Lane 12: PC-12 cell lysate (20 µg/Lane) Lane 13: Mouse kidney tissue lysate (40 µg/Lane) Lane 14: Rat kidney tissue lysate (40 µg/Lane)
Predicted band size: 34 kDa Observed band size: 34 kDa
Exposure time: 40 seconds;
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 (HA721509) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
Immunofluorescence analysis of paraffin-embedded mouse small intestine tissue labeling Syntaxin 4 with Rabbit anti-Syntaxin 4 antibody (HA721509) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA721509, green) at 1/200 dilution overnight at 4 ℃, washed with PBS.
Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
☑ Knockdown (KD)
Western blot analysis of Syntaxin 4 on different lysates with Rabbit anti-Syntaxin 4 antibody (HA721509) at 1/5,000 dilution.
Lane 1: HAP1-parental cell lysate Lane 2: HAP1-Syntaxin 4 KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 34 kDa Observed band size: 34 kDa
Exposure time: 3 minutes; 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 (HA721509) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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 prostate tissue with Rabbit anti-Syntaxin 4 antibody (HA721509) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721509) 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 human skin tissue with Rabbit anti-Syntaxin 4 antibody (HA721509) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721509) 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 human small intestine tissue with Rabbit anti-Syntaxin 4 antibody (HA721509) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721509) 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 small intestine tissue with Rabbit anti-Syntaxin 4 antibody (HA721509) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721509) 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 kidney tissue with Rabbit anti-Syntaxin 4 antibody (HA721509) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA721509) 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.
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