Samples of interest were lysed in RIPA buffer (Cell Signaling 9806) containing protease inhibitor cocktail (Roche 11836170001), 1 mM PMSF, and phosphatase inhibitor (Pierce A32957), unless noted otherwise. Protein lysate concentrations were normalized using Pierce BCA (Thermo 23225), and then reduced and denatured in sample buffer (Thermo 39000). Lysates were separated on Bio-Rad 4%–20% Mini-PROTEAN TGX precast gels, transferred via Bio-Rad Trans-Blot Turbo onto PVDF membranes (Bio-Rad 1704272), blocked in TBS-T (TBS with 0.1% Tween-20) containing 5% BSA, and probed overnight at 4°C with primary antibody. The next day, blots were washed in TBS-T and probed with HRP-linked secondary antibody (Cell Signaling 7076; 7074; Streptavidin-HRP 3999S) for 1 hour. Signal detection was performed using ECL substrate (Thermo 34577; 34095) and a Bio-Rad ChemiDoc MP imaging system. Blot images were digitally processed and analyzed in ImageJ. The primary antibodies used were as follows: anti-actinin (isoforms 1–4) (Cell Signaling 6487), anti-HA (Cell Signaling 14793), anti-IGF2BP2 (MBL RN008P), anti-PBCBP1 (Abcam 74793), anti-PCBP2 (MBL RN250P), anti-SERBP1 (Abcam 55993), anti-TCAP (BD 612328), anti-FLAG (Cell Signaling 2368), anti-GAPDH (Cell Signaling 2118) and anti-MYOM (mMaC myomesin B4 was deposited to the DSHB by Perriard, J.-C.).
Anti serbp1
Manufactured by Abcam
Sourced in United States
Anti-SERBP1 is a protein-specific antibody that recognizes the SERBP1 (Serpine1 mRNA Binding Protein 1) protein. SERBP1 is involved in the regulation of mRNA stability and translation. The antibody can be used for applications such as western blotting, immunohistochemistry, and immunofluorescence.
Automatically generated - may contain errors
Lab products found in correlation
Mini protean tgx precast gel, by Bio-Rad (2 mentions)
Pierce bca, by Thermo Fisher Scientific (2 mentions)
Trans blot turbo, by Bio-Rad (2 mentions)
Pvdf membrane, by Bio-Rad (2 mentions)
Sample buffer, by Thermo Fisher Scientific (2 mentions)
Ripa buffer, by Cell Signaling Technology (2 mentions)
Anti ha, by Cell Signaling Technology (2 mentions)
Anti gapdh, by Cell Signaling Technology (2 mentions)
Protease inhibitor cocktail, by Roche (2 mentions)
Chemidoc mp imaging system, by Bio-Rad (2 mentions)
Anti pbcbp1, by Abcam (2 mentions)
Anti tcap, by BD (2 mentions)
Anti myom, by Cell Signaling Technology (2 mentions)
Anti flag, by Cell Signaling Technology (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Gapdh, by Abcam (1 mentions)
4 protocols using anti serbp1
1
Western Blot Analysis of Protein Targets
2
Western Blot Analysis of Protein Targets
3
Western Blot Analysis of Actin and SERBP1
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Cell lysates were analyzed by Western blotting using the following primary antibodies: Anti-actin rabbit polyclonal (Abcam), anti-SERBP1 (Abcam) followed by the appropriate horseradish peroxidase (HRP)-conjugated secondary antibody, and analyzed by chemiluminescence.
4
Transfection of miR-362-3p in cell lines
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The anti-SERBP1 and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) primary antibodies were purchased from Abcam (Cambridge, MA, USA) and CST (Cell Signaling Technology, USA), respectively. The miR-362-3p mimic, miR-362-3p inhibitor, and negative control (NC) (scrambled sequence) were synthesized by and purchased from Shenggong (Shanghai, China). The sequences are presented in Table 1 . All these molecules were transfected into cells using Lipofectamine 2000 (Invitrogen, USA) according to the manufacturer’s protocol.
The sequence of miRNA
| miRNA | Sequence |
|---|---|
| miR-362-3p | 5’-AACACACCUAUUCAAGGAUUCA-3’ |
| miR-362-3p mimics | 5’-AACACACCUAUUCAAGGAUUCA-3’ |
| miR-362-3p inhibitor | 5’-UGAAUCCUUGAAUAGGUGUGUU-3’ |
| miRNA NC (Scramble) | 5’-UUCUCCGAACGUGUCACGUTT-3’ |
NC, negative control
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