Rabbit anti human gapdh

MGC80-3 and AGS cells were harvested at the 5^th day post infection. Then cells were lysed in 2× SDS Sample Buffer (100 mM Tris-HCl, pH = 6.8, 10 mM EDTA, 4% SDS, 10% Glycine). The concentration of protein in the cell lysate was determined using the BCA protein assay kit (Pierce Biotechnology, Cat no. 23235). Totally, 30 μg of protein was loaded on per lane, followed by separation using SDS-PAGE, and transfer to PVDF membrane. Then the membrane was subsequently incubated with primary antibodies, including rabbit anti-human-GGCT (Sigma, Cat no. HPA020735, 1:500), rabbit anti-human-GAPDH (Proteintech Group, Inc., Cat no. 10494-1-AP, 1:60000), overnight at 4 °C, which was followed by incubation with secondary antibody, horseradish peroxidase-conjugated anti-rabbit IgG (Santa Cruz, Cat no.SC-2054, 1:5000), for 1 h at room temperature. Blots were visualized using ECL Test Kit (Amersiam, Cat no. RPN2132). GAPDH was used as the internal control. Density analysis was carried out using Quantity One software (BioRad).

Western blot was conducted as described previously.¹⁹ The primary antibodies were rabbit anti‐human p75NTR (1:1000; Abcam), rabbit anti‐human alkaline phosphatase (ALP) (1:500; Abcam), rabbit anti‐human RUNX family transcription factor 2 (RUNX2) (1:1000; Abcam) and rabbit anti‐human α1 integrin (ITGA1) (1:500; Abcam). Rabbit anti‐human GAPDH (1:10 000; Proteintech, Wuhan, China) was used as an internal standard.