Total proteins were resolved using 8–10% SDS-PAGE, transferred onto PVDF membranes, and blocked using 3% BSA dissolved in TBST (Tris-buffered saline, 0.1% Tween 20) for 2 h at room temperature. The membranes were incubated with primary antibodies against WTAP (Abcam; catalog ab195380; 1:1000), METTL3 (Abcam; catalog ab195352; 1:1000), METTL14 (Abcam; catalog ab220030; 1:500), KIAA1429 (Proteintech, catalog 25712-1-AP; 1:500), Cleaved PARP (Abcam; catalog ab32064; 1:1000), Cleaved Caspase-3 (Abcam; catalog ab2302; 1:500), p-PERK (Biorbyt; catalog orb191598; 1:1000), PERK (Abcam; catalog ab65142; 1:500), p-eIF2α (Cell Signaling Technology; catalog #9721; 1:1000), eIF2α (Cell Signaling Technology; catalog #5324; 1:1000), ATF4 (Cell Signaling Technology; catalog #11815; 1:1000),CHOP (Cell Signaling Technology; catalog #2895; 1:1000), EIF3A (Abcam; catalog ab128996; 1:1000), and GAPDH (Abcam; catalog ab9485; 1:1000), then incubated with secondary antibodies (Beyotime; catalog A0208, catalog A0216)Protein bands were quantified using Image J.
Ab220030
Manufactured by Proteintech
Ab220030 is an anti-human PCSK9 antibody produced by Proteintech. It is a recombinant polyclonal antibody designed for research applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab32064, by Abcam (1 mentions)
Eif2α, by Cell Signaling Technology (1 mentions)
Ab195352, by Abcam (1 mentions)
Ab195380, by Abcam (1 mentions)
Ab128996, by Abcam (1 mentions)
Ab9485, by Beyotime (1 mentions)
P eif2α, by Cell Signaling Technology (1 mentions)
A0208, by Beyotime (1 mentions)
Ripa lysis buffer, by Thermo Fisher Scientific (1 mentions)
Goat anti rabbit igg h l hrp, by Beyotime (1 mentions)
Ecl system, by Thermo Fisher Scientific (1 mentions)
Goat anti mouse igg h l hrp, by Beyotime (1 mentions)
2 protocols using ab220030
1
Quantitative Protein Analysis for M6A Modulators
2
Quantitative Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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RIPA lysis buffer (Thermo) was used to extract total protein products from the cells. Next, proteins were separated on 8% SDS‐PAGE and transferred to PVDF membranes. The membranes were blocked with skimmed milk for 3 h at 37°C and then incubated with primary antibody overnight at 4°C. Finally, the membranes were incubated with secondary antibody for 2 h at 37°C. The ECL System (Thermo) was used to develop the films, and then quantified with ImageJ software. The primary antibodies were as follows: anti‐METTL14 (Abcam, #ab220030, 1:1000) and anti‐GAPDH (Proteintech, #60004‐1‐Ig, 1:10000). The secondary antibodies were as: goat anti‐rabbit IgG H&L (HRP) (Beyotime, #A0208, 1:1000) and goat anti‐mouse IgG H&L (HRP) (Beyotime, #A0216, 1:1000).
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