Hrp conjugated anti mouse igg sc 2005

Cells were harvested and resuspended in SDS buffer (Beyotime, Shanghai, China) for preparation of total protein extracts. Briefly, total protein extract was separated by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto an Immobilon-P transfer polyvinylidene fluoride membrane (Millipore, MA, USA). The membrane was incubated at room temperature in 5% bovine serum albumin (BSA) prepared with TBS-T buffer for 2 h. The membrane was incubated with primary antibody diluted 1∶1,000 with 1% BSA at 4°C overnight. On the following day, the membrane was incubated with secondary horseradish peroxidase (HRP)-conjugated antibody diluted 1∶5,000 with 1% BSA at 4°C for 6 h. The membrane was visualized after incubation in HRP substrate (BeyoECL plus A/B, Beyotime, Shanghai, China). The antibodies used in this study were anti-cyclin D1 (sc-8396, Santa Cruz, CA, USA), anti-CDK4 (sc-260, Santa Cruz, CA, USA), anti-β-actin (A2066, Sigma, MO, USA), HRP-conjugated anti-mouse IgG (sc-2005, Santa Cruz, CA, USA), and HRP-conjugated anti-rabbit IgG (sc-2004, Santa Cruz, CA, USA).

The following antibodies were used for immunoblotting: mouse anti-TurboGFP (TA150041) [Origene]; rabbit anti-TDP-43 (10782-2-AP) [Proteintech]; mouse anti-Lamin A/C (05-714) [EMD Millipore]; rabbit anti-beta Actin (ab16039) [Abcam]; rabbit anti-NF-κB p65 (Ser536) (3033), rabbit anti-NF-κB (8242) and HRP-conjugated anti-α-tubulin (9099) [Cell Signaling Technology]; and rabbit anti-PTP1B (sc-14021), HRP-conjugated anti-rabbit IgG (sc-2004) and HRP-conjugated anti-mouse IgG (sc-2005) [Santa Cruz]. The following antibodies were used for immunocytochemistry (ICC): rabbit anti-MAP2 (1:500; AB5622) and TRITC-conjugated phalloidin (1:500; 90228) [Millipore]. The following antibody was used for immunohistochemistry (IHC): FITC-conjugated anti-HRP (1:150; 123-095-021) [Jackson ImmunoResearch Laboratories]. The following antibodies were used for neutralizing target proteins: rabbit anti-IL-1β (ab9722) [Abcam]; rat anti-TNF-α (14-7321-81) [Invitrogen]; rabbit anti-IgG (2729) and mouse anti-IgG (5415) [Cell Signaling Technology]; and mouse anti-IL-6 (sc-57315) and rat anti-IgG (sc-2026) [Santa Cruz].