The slices on slides were subjected to dewaxing and rehydration. Subsequently, permeabilization was performed using 1% Triton X-100 (#T8200; Solarbio) and blocking was performed with 10% horse serum. Next, the sections were incubated overnight at 4°C with the following primary antibodies: Ptgs2 (1:200, ab283574; Abcam, Cambridge, UK), Gpx4 (1:200, ab125066; Abcam), hepcidin (1:200, AF7003; Affinity, Liyang, China), IGF-1R (1:200, AF6125; Affinity), P-IGF-1R (1:200, AF3123; Affinity), P-AKT (1:200, AF0016; Affinity), ferritin (1:100, ab287968; Abcam) and TfR1 (1:2000, ab269513; Abcam). Then, the HRP-conjugated secondary antibody (PV-6000; ZSGB-BIO, Beijing, China) was used for incubation at 37°C for 20 min. Reaction products were visualized by using 3,3′-diaminobenzidine (DAB). The images were captured with the PA53 biological microscope.
Ab283574
Manufactured by Abcam
Sourced in United Kingdom
Ab283574 is a lab equipment product from Abcam. It is designed for a core function, the details of which are not available for an unbiased and factual description without extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
Ab269513, by Abcam (1 mentions)
Ab287968, by Abcam (1 mentions)
Ab125066, by Abcam (1 mentions)
Pv 6000, by ZSGB-BIO (1 mentions)
Af7003, by Affinity Biosciences (1 mentions)
Af0016, by Affinity Biosciences (1 mentions)
Af6125, by Affinity Biosciences (1 mentions)
T8200, by Solarbio (1 mentions)
Ab283655, by Abcam (1 mentions)
Ab86299, by Proteintech (1 mentions)
Cw0027s, by CWBIO (1 mentions)
Ab16502, by Abcam (1 mentions)
2 protocols using ab283574
1
Immunohistochemical Analysis of Oxidative Stress Markers
2
Western Blot Analysis of Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The protein was then premixed with SDS-PAGE (CW0027S, CWBIO) in a 4:1 ratio and heated at 100 ℃ for 10 min. The protein samples were separated and transferred to polyvinylidene fluoride (PVDF) membrane. Then, the PVDF membrane was blocked with 5% nonfat milk for 1 h and incubated overnight with a primary antibody. Subsequently, the PVDF membranes were incubated with corresponding secondary antibodies at room temperature for 1 h and ECL Kit Chemiluminescence (S6008M, US EVERBRIGHT) was applied to visualize the immunoblots. Finally, ImageJ software was used for protein analysis.
The primary antibodies used in Western blot are as follows: anti-mas1 (1:1000, Proteintech, 20080–1-ap), anti-IL-1β (1:2000, Proteintech, 26,048–1-AP), anti-TNF-α (1:1000, Proteintech, 60291–1-Ig), anti iNOS (1:1000, Abcam, ab283655), anti-Arg-1 (1:1000, Abcam, ab283574), anti-p65(1:1000, Abcam, ab16502), anti-p65 (phospho S536) (1:1000, Abcam, ab86299), anti-TLR4(1:1000, Proteintech, 19811–1-AP), anti-IκB(1:2000, Proteintech, 10268–1-AP), anti-GAPDH (1:1000, Abcam, ab18602), anti-βactin (1:20000, Proteintech, 66009–1-Ig), anti-vinculin (1:10000, Proteintech, 66305–1-Ig).
The primary antibodies used in Western blot are as follows: anti-mas1 (1:1000, Proteintech, 20080–1-ap), anti-IL-1β (1:2000, Proteintech, 26,048–1-AP), anti-TNF-α (1:1000, Proteintech, 60291–1-Ig), anti iNOS (1:1000, Abcam, ab283655), anti-Arg-1 (1:1000, Abcam, ab283574), anti-p65(1:1000, Abcam, ab16502), anti-p65 (phospho S536) (1:1000, Abcam, ab86299), anti-TLR4(1:1000, Proteintech, 19811–1-AP), anti-IκB(1:2000, Proteintech, 10268–1-AP), anti-GAPDH (1:1000, Abcam, ab18602), anti-βactin (1:20000, Proteintech, 66009–1-Ig), anti-vinculin (1:10000, Proteintech, 66305–1-Ig).
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