Capsaicin (CAP) and Sorafenib were purchased to Sigma (St. Louis, MO, USA). Primary antibodies anti-caspase-9, anti-PARP, anti-AFP, anti-pAkt-ser473, p-mTOR-ser2448, p-AMPKα1-thr172, p-ACC-ser79 and the antibodies against the corresponding total forms were obtained from Cell Signaling Technology (Danvers, MA, USA). The anti-caspase-3 antibody was from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). (Peroxidase labeled secondary anti-mouse IgG was from Sigma (St. Louis, MO, USA) and anti-rabbit IgG was from Calbiochem (San Diego, CA, USA).
Anti afp
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-AFP is a laboratory reagent produced by Cell Signaling Technology. Its core function is to detect and quantify alpha-fetoprotein (AFP) in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Sorafenib, by Merck Group (1 mentions)
P mtor ser2448, by Cell Signaling Technology (1 mentions)
Peroxidase labeled secondary anti mouse igg, by Merck Group (1 mentions)
P ampkα1 thr172, by Cell Signaling Technology (1 mentions)
P acc ser79, by Cell Signaling Technology (1 mentions)
Capsaicin cap, by Merck Group (1 mentions)
Anti caspase 3 antibody, by Santa Cruz Biotechnology (1 mentions)
Anti p akt ser473, by Cell Signaling Technology (1 mentions)
Anti caspase 9, by Cell Signaling Technology (1 mentions)
Anti parp, by Cell Signaling Technology (1 mentions)
Anti cyclin b1, by Abcam (1 mentions)
Anti 53bp1, by Cell Signaling Technology (1 mentions)
Anti cd45, by Agilent Technologies (1 mentions)
Anti phospho stat3, by Abcam (1 mentions)
Anti ck19, by Agilent Technologies (1 mentions)
Anti hnf4, by Santa Cruz Biotechnology (1 mentions)
Anti pr set7, by Cell Signaling Technology (1 mentions)
Anti ki67, by Abcam (1 mentions)
Anti stat3, by Abcam (1 mentions)
Anti sma, by Abcam (1 mentions)
6 protocols using anti afp
1
Apoptosis and Proliferation Signaling
2
Comprehensive Antibody Resource for Liver Research
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The antibodies used in this study were from the following: Santa Cruz Biotechnology: anti-HNF4 (sc-8987), anti-AFP (sc-8108), anti-Pr-Set7 (sc-135009), anti-53BP1 (sc-22760) and anti-FGF7 (SC-27127); Cell Signaling Technology: anti-cyclin B1 (#4138), anti-Stat3 (#9132), anti-phospho-Stat3 (#9145) and anti-phospho-histone H2A.X (#9718); Abcam: anti-Ki67 (ab15580), anti-histone H4 mono methyl K20 (ab9051), anti-histone H4 tri methyl K20 (ab9053) and anti-Pr-Set7 (ab3798); Bethyl Laboratories, anti-Alb (A90–234); AbD Serotec, anti-F4/80 (MCA497); Merck-Millipore, anti-Sox9 (AB5535) and anti-Prominin-CD133 (MAB4310); Biolegend, anti-CD45 (#103101); and DAKO, anti-CK19 (#A0575). The A6 antibody was obtained from Valentina Factor (NIH).
3
Teratoma Formation Assay for iPSCs
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When the iPSCs reached 80% confluence 6 cm dishes, they were harvested by collagenase IV treatment, collected into tubes and injected below the capsule of the testes of 6- to 12-week-old SCID mice. After 9 to 12 weeks of injection, the teratomas were collected and fixed with 4% paraformaldehyde. The paraffin-embedded tissue was sectioned and stained with hematoxylin and eosin (H&E). The paraffin embedded sections were deparaffinized and rehydrated in Target Retrieval Solution (DaKo). Sections were blocked with 3% fetal bovine serum for 5 mins and incubated with the primary antibodies for 30 mins at room temperature. The primary antibodies used in this assay were anti-AFP (1:100; Cell Signaling), anti-GFAP (1:300; Cell Signaling), anti-SMA (1:100; Abcam).
4
Western Blot Analysis of Cell Lines
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For Western blot analysis whole-cell extracts were prepared from mouse (AML12, BNL Cl.2), and human (HepG2) cell lines. Certain proteins were isolated from supernatant collected from HepG2 cells. The nuclear protein isolation was performed using Abcam Nuclear Extraction Kit (ab113474). Proteins were separated on a 10% or 4% to 20% gradient Mini-PROTEAN TGX Precast Protein Gels (Bio-Rad). The anti–trefoil factor 3 (TFF3) antibody (Thermo Fisher Scientific PA5–21081) was used at 1:500, the anti–insulin-like growth factor receptor 1 (IGFR1; phospho Y1161) antibody (Abcam AB_731544) was used at 1:100, the anti–insulin-like growth factor binding protein 5 (IGFBP5) antibody (Thermo Fisher Scientific PA5–37369) was used at 1:1,000, anti-Histone H3 antibody (Abcam AB_302613) was used at 1:1,000 and the anti-AFP, anti-tubulin, anti-GFP, or β-actin antibody (Cell Signaling Technology, #2137; Cell Signaling Technology, #9099; Cell Signaling Technology, #2956; Cell Signaling Technology, #4967) were used at a 1:2,000 dilution.
5
Antibody Sources for Protein Analysis
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Specific antibodies were purchased from the following commercial sources: Anti-AFP, anti-ALB, anti-CD44, anti-Evi1, anti-flag (mouse), anti-HA, anti-HNF4α, anti-H3, anti-H3K36me3, anti-Myc, anti-OCT4A, anti-P300, anti-PPM1A (rabbit), anti-pSmad2 (S465/467), anti-pSmad2 (S245/250/255), anti-pSmad3 (s423/425), anti-Smad2, anti-Smad3 (rabbit), anti-SnoN, anti-Sox2, anti-TAT, and normal rabbit IgG from Cell Signaling Technology (Danvers, MA); anti-PPM1A (mouse), anti-SC35, and anti-SETD2 were from Abcam (Cambridge, MA); Anti-Smad4 and normal mouse IgG were from Santa Cruz Biotechnology (Santa Cruz, CA); Anti-β-actin, and anti-flag (rabbit) from Sigma-Aldrich (St. Louis, MO); Alexa594 goat anti-mouse IgG from Life Technology (Carlsbad, CA); Dylight488 goat anti-rabbit IgG from Vector Labs (Burlingame, CA).
6
Western Blot Analysis of Protein Markers
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The total protein lysate of cells was prepared in radioimmunoprecipitation assay buffer containing 1 mM phenylmethylsulfonyl fluoride (Beyotime, Shanghai, China). A bicinchoninic acid protein assay kit (Beyotime) was used to determine the protein concentrations. Equal amounts (20 μg) of proteins were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto a polyvinylidene difluoride membrane (Millipore, Billerica, MA, USA). Subsequently, the membrane was incubated in 5% nonfat milk in Tris-buffered saline (TBS) for 1 h at room temperature. The following primary antibodies were incubated with the membrane at 4°C overnight: anti-CXCR4 (cat:#ab181020, 1 : 1,000) from Abcam (Cambridge, MA, USA), anti-AFP (cat:#4448, 1 : 1,000), anti-FOXM1 (cat:#20459, 1 : 1,000), phosphor-FOXM1 (cat:#14170, 1 : 1,000), and anti-p65 (cat:#3033, 1 : 1,000) from Cell Signaling Technology (Danvers, MA, USA), as well as anti-β-actin (cat:#66009-1-Ig, 1 : 5,000) from Proteintech. After washing with 0.5% Tween-20 in TBS, the membrane was incubated with horseradish peroxidase-conjugated secondary antibody (Absin, Shanghai, China) for 1 h at room temperature. Protein signals were developed with enhanced chemiluminescence.
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