Protein concentration was determined by the Bradford assay. 10% SDS-PAGE was used for protein separation. Protein bands were transferred onto a nitrocellulose (NC) membrane (PALL, USA), blocked with 5% (w/v) skim milk (BD, USA) in TBST (50 mM Tris, 100 mM NaCl and 0.1% Tween-20, pH 7.5) and incubated with primary antibodies FAM129A (1:1000, Proteintech, USA), GAPDH (1:2000, Proteintech, USA), TIMP1(1:1000, Proteintech, USA), MMP2 (1:500, Proteintech, USA), MMP9 (1:500, Proteintech, USA), MDM2 (1:500, Proteintech, USA), P53 (1:1000, Proteintech, USA), Bcl2 (1:1000, Wanleibio, China) and Bax (1:1500, Proteintech, USA) at 4 °C with shaking at 100 rpm overnight. The NC membrane was washed well with TBST for 3 × 10 min, incubated with HRP-conjugated Affinipure Goat Anti-Rabbit IgG (1:2000, Proteintech, USA) for 2 h at RT, and then washed again with TBST for 3 × 10 min. Protein bands were visualized by ECL and quantified by the Bio-Rad ChemiDoc™ MP system (Bio-Rad, USA).
Fam129a
Manufactured by Proteintech
Sourced in United States
FAM129A is a laboratory reagent product offered by Proteintech. It is a protein that can be used in various research applications. The core function of FAM129A is to serve as a tool for scientific investigation, without making claims about its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Timp1, by Proteintech (2 mentions)
Skim milk, by BD (1 mentions)
Chemidoc mp system, by Bio-Rad (1 mentions)
Affinipure goat anti rabbit igg, by Proteintech (1 mentions)
Gapdh, by Proteintech (1 mentions)
Bcl 2, by Wanlei (1 mentions)
Bcl2 associated x (bax), by Proteintech (1 mentions)
Bx63 microscope, by Olympus (1 mentions)
Biotin streptavidin horseradish peroxidase hrp detection system, by ZSGB-BIO (1 mentions)
2 protocols using fam129a
1
Protein Expression Analysis Protocol
2
Evaluating FAM129A, MMP9, and TIMP-1 in HCC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Expression levels of FAM129A (Proteintech, USA), MMP9 (Proteintech, USA) and TIMP-1 (Proteintech, USA) were determined by IHC assay in HCC tissue arrays. A biotin-streptavidin horseradish peroxidase (HRP) detection system (ZSGB-BIO, China) was used to treat samples according to the manufacturer’s guidelines and a DAB (diaminobenzidine) kit (ZSGB-BIO, China) was used to envision images using a BX63 microscope (Olympus, Japan). Score I represents IHC immunoreactive intensity and was classified into four ranks, 0 (negative), 1 (weak), 2 (moderate) and 3 (strong), whereas Score II signifies the observed positively stained cells, and the DAB staining level of each sample was categorized as 0 (none), 1 (1–10% cells per field), 2 (10–50%), 3 (51–75%) and 4 (>76%). The IHC immunoreactivity degree is the product of Score I and Score II and ranges from 0 to 12. The scores were considered in following way: 0–2 as negative (−), 3–5 as weak (+), 6–8 as moderate (++), and 9–12 as strong (+++). Scoring of IHC assays was done independently by two experienced pathologists.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!