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Ab109451

Manufactured by Proteintech

Ab109451 is a primary antibody that recognizes the human Transferrin Receptor protein. It is suitable for use in various immunoassay applications.

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3 protocols using ab109451

1

Immunoblot Analysis of PRMT5 Interactome

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For immunoblot analysis: Antibodies purchased from Cell Signaling include: PRMT5 Ab (2252 s, 1:1000), Total Rb (4H1) mAb (9309 s, 1:1000), pRB/s807/811 (D20B12) mAb (8516 s, 1:1000), β-actin (13E5) mAb (4970 s, 1:5000), Vinculin (E1E9V) mAb (13901 s, 1:2000), SDMA Ab (13222 s, 1:1000), MEP50 Ab (2823 s, 1:1000), PRMT5 (D5P2T) mAb (79998 s, 1:1000), and ORC2 (3G6) mAb (4736 S, 1:1000); Antibodies purchased from Santa Cruz Biotechnology include: ERα (F-10) mAb (sc-8002, 1:1000) and POLE (D-10) mAb (sc-390785, 1:1000); Antibodies purchased from Abcam: PRMT5 (EPR5772) mAb (ab109451, 1:1000) and GINS1 (EPR13359) mAb (ab181112, 1:1000); FUS Ab was purchased from Proteintech (11570-1-AP, 1:1000); Total Pol II (4H8) mAb was purchased from MilliporeSigma (05-623, 1:1000); APC7 Ab was purchased from Bethyl Laboratories (A302-551A, 1:1000). For Co-IP: PRMT5 mAB (EPR5772) was purchased from Abcam (ab109451, 1:250); FUS mAB (4H11) was purchased from Santa Cruz Biotechnology (sc-47711, 1:100). For ChIP-seq: pSer2 Pol II Ab was purchased from Abcam (ab5095, 1:40). For IHC, SDMA Ab was purchased from Cell Signaling (13222 s, 1:600); ERα mAb (F-10) was purchased from Santa Cruz Biotechnology (sc-8002, 1:800); Ki67 (MIB-1) mAb was purchased from Agilent (IR62661-2, ready to use).
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2

Protein Interactions in Hepatocellular Carcinoma

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These experiments were performed as described previously.
11 (link),
12 (link) For Co‐IP, HCC cell lines including MHCC‐97H, Huh7‐SMYD4, Huh7 pre‐transfected with plasmids of Flag‐labeled different PRMT5 regions, and HA‐labeled SMYD4 were used. The cell protein lysates were mixed with antibodies of PRMT5 (Abcam, ab109451), SMYD4 (Proteintech, 17594‐1‐AP), HA (Abcam, ab9110), or Flag (Cell Signaling Technology, #14793) according to each experiment. Silver staining was performed using a silver stain kit SilverQuest™ (ThermoFisher Scientific) according to the manufacturer's instructions. LS‐MS detection and analysis were performed by BGI Company.
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3

Protein Expression Profiling in Colorectal Cancer Cell Lines

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Whole-cell lysates of HCT116 and SW480 cells were extracted using lysis buffer, including 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.1% SDS, 1% sodium deoxycholate and 1 mM phenylmethylsulfonyl fluoride (PMSF). Cell lysates were centrifuged at a speed of 14000 rpm for 15 min, then mixed with 5×loading buffer and denatured at 100 °C for 5 min. Aliquots of supernatant protein (40 ~ 50 μg) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and electrophoretically transferred to a polyvinylidene fluoride (PVDF) membrane (Millipore). The PVDF membranes were then blocked for 1 h with 5% (w/v) non-fat milk at room temperature. After incubation with anti-PRMT5 (Abcam, ab109451), anti-EZH2 (ProteinTech, 21800-1-AP), anti-CDKN2B (Abcam, ab53034) or anti-GAPDH (ProteinTech, 60004) overnight at 4°C, the PVDF membranes were washed with PBST and incubated with corresponding horseradish peroxidase (HRP)-conjugated secondary antibody (Sigma-Aldrich) at room temperature. Subsequently, the immunoreactive bands were visualized using ECL detection reagent (Thermo Scientific).
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