We randomly selected 84, 73 and 62 patients from the followed 588 patients according to BMI levels, respectively. The breast cancer tissues and adjacent normal tissues were stained with HPGD protein polyclonal antibody (1: 200, A6926; ABclonal, USA). The protocol for IHC has been described in detail in previous Publications [41 (link)]. HPGD protein immunohistochemical score was independently evaluated in a blinded fashion based on the staining intensity and the percentage of positive cells, resulting in scores of 0–7. Scores of 0–4 were considered as low HPGD protein expression and scores of 5–7 were considered as high HPGD protein expression [17 (link)].
A6926
Manufactured by ABclonal
Sourced in United States, China
A6926 is a laboratory centrifuge designed for a variety of applications. It has a rotor capacity of up to 4 x 100 mL. The centrifuge can achieve a maximum speed of 6,000 rpm and a maximum relative centrifugal force of 4,500 x g.
Automatically generated - may contain errors
2 protocols using a6926
1
HPGD Protein Expression in Breast Cancer
2
Protein Extraction and Western Blot Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total protein was extracted using RIPA lysate and PMSF. The protein concentration was determined by BCA Protein Assay Kit (Solarbio, China). Polyacrylamide gel is consisted of 5% stacking gel and 12% and 14% separating gel. SDS-PAGE was performed to separate the target protein. Then the proteins were transferred to PVDF membrane, which was subsequently blocked with blocking solution (Solarbio, China). Incubation of primary antibody was conducted at 4℃ overnight and secondary antibody incubation was performed at 37℃ for 1 h. After incubation of antibodies, the protein bands were detected by ECL-based chemiluminescence reaction. The optical density of the target band was analyzed by Gel-Pro-Analyzer software (USA). The antibodies were as follows: HPGD antibody (1:500 dilution, A6926, Abclonal, China); Nr4a1 antibody (1:2000 dilution, 12235-1-AP, proteintech, China); GAPDH (1:10000 dilution, 60004-1-Ig, proteintech, China); Goat anti-rabbit IgG-HRP (1:3000 dilution, SE134, Solarbio, China); and Goat anti-mouse IgG-HRP (1:3000 dilution, SE131, Solarbio, China).
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