Cells or tissues were lysed with RIPA lysis (Beyotime, Shanghai, China) to collect total protein. The protein concentration was determined by BCA protein assay kit (Beyotime, Haimen, China). The protein was separated using 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membrane. After being blocked with 5% skim milk, the membrane was incubated with primary antibodies: anti-vimentin (1:1000, # 5741; CST), anti-E-cadherin (1:1000, # 3195; CST), anti-RAB11A (1:500, ab128913; Abcam), and anti-β-actin (1:2000, ab179467; Abcam). Subsequently, the membrane was incubated with the secondary antibody: goat anti-rabbit (1:5000, ab6721; Abcam) or goat anti-mouse (1: 5000, ab6728; Abcam). Band development was carried out utilizing ECL kit (Amersham Pharmacia Biotech, Little Chalfont, United Kingdom) and photographs were taken using ImageQuant LAS 4000 micro biomolecular imager (GE Healthcare).
Ab128913
Manufactured by Abcam
Ab128913 is a high-quality antibody product offered by Abcam. This antibody is designed for use in research applications. No further details about its core function or intended use are provided in order to maintain an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Ab6721, by Abcam (1 mentions)
Ab179467, by Abcam (1 mentions)
Ripa lysis, by Beyotime (1 mentions)
Ecl kit, by Cytiva (1 mentions)
Imagequant las 4000, by GE Healthcare (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Ab6728, by Abcam (1 mentions)
Ab80864, by Abcam (1 mentions)
Ab2900, by Abcam (1 mentions)
A10037, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 568 donkey anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
A10042, by Thermo Fisher Scientific (1 mentions)
2 protocols using ab128913
1
Western Blot Analysis of Epithelial and Mesenchymal Markers
2
Measuring Autophagy Flux in HeLa Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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HeLa cells were fixed using 4% paraformaldehyde or ice-cold methanol for 10 min. The cells were then stained using antibodies against giantin (ab80864, Abcam), RAB11 (ab128913, Abcam), EEA1 (ab2900, Abcam), and LC3B (no. 2775, Cell signaling Technology) overnight at 4°C. Subsequently, cells were incubated with donkey anti-mouse IgG–Alexa Fluor 568 (A10037, Invitrogen) or donkey anti-rabbit IgG–Alexa Fluor 568 (A10042, Invitrogen). To determine autophagic flux, the colored puncta in cells expressing mRFP-GFP-GABARAPL2 and mRFP-GFP-LC3B were counted with the ImageJ macro “Red and Green Puncta Colocalization Macro.”
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