Total proteins from flight muscles of WT and ACP-/- locusts were extracted using TRIzol reagent, as previously described (Hou et al., 2019 (link)). The protein extracts (50 μg) were electrophoresed on 4–20% Biofuraw precast gels (Tanon, China) and then transferred to polyvinylidene difluoride (PVDF) membranes (Millipore). The membrane was incubated with polyclonal antibody against target protein (anti-FABP, developed by ABclone, China, 1:5000; anti-CPT2, Abcam, 1:1000; anti-ACDM, Abcam, 1:1000). Goat anti-rabbit IgG (EASYBIO, 1:5000) was used as the secondary antibody. Polyclonal antibody against tubulin was used as an internal control. Protein bands were detected by chemiluminescence (ECL kit, Thermo Scientific).
Goat anti rabbit igg
Manufactured by EASYBIO
Sourced in China
Goat anti-rabbit IgG is a secondary antibody that binds to rabbit immunoglobulin G (IgG). It is commonly used in various immunoassay techniques, such as Western blotting, ELISA, and immunohistochemistry, to detect and quantify the presence of rabbit-derived primary antibodies.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti mouse igg, by EASYBIO (3 mentions)
Ecl kit, by Thermo Fisher Scientific (2 mentions)
Anti cpt2, by Abcam (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Nitrocellulose membrane, by GE Healthcare (1 mentions)
Goat anti mouse igg horseradish peroxidase conjugate, by Bio-Rad (1 mentions)
Biomolecular imager, by Azure Biosystems (1 mentions)
Be0102, by EASYBIO (1 mentions)
Be0101, by EASYBIO (1 mentions)
Be0031, by EASYBIO (1 mentions)
As09532a, by Agrisera (1 mentions)
Phosphatase inhibitor, by Applygen (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Cleaved caspase 8, by Immunoway (1 mentions)
Caspase 8, by Immunoway (1 mentions)
Cleaved caspase3, by Immunoway (1 mentions)
Cox 2, by ABclonal (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Interleukin 6 (il 6), by ABclonal (1 mentions)
Caspase3, by Immunoway (1 mentions)
5 protocols using goat anti rabbit igg
1
Protein Extraction and Western Blot Analysis of Locust Muscle
2
Protein Expression Analysis of Beetle Trachea
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total proteins were extracted from thoracic tracheal tubes of each beetle in different treatments using TRIzol reagent according to the manufacturer’s protocol and stored at –80°C until used in experiments. The protein extracts (100 mg) were electrophoresed on 4–12% SDS-PAGE precast gels (EASYBIO, China) and then transferred to 0.22 μm polyvinylidene difluoride (PVDF) membranes (Millipore). The membrane was incubated with polyclonal antibody against target protein (anti-Muc91C, developed by BGI, China, 1:4000). Goat anti-rabbit IgG (EASYBIO, 1:5000) was used as the secondary antibody. Monoclonal antibody against Histone H3 (EASYBIO, 1:2000) was used as an internal control. Goat anti-mouse IgG (EASYBIO, 1:5000) was used as the secondary antibody. Protein bands were detected by SuperSignal West Atto Ultimate Sensitivity Substrate (Invitrogen, A38554).
3
Western Blot Analysis of Recombinant Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total proteins were isolated from N. benthamiana leaf tissues in extraction buffer (100 mM Tris-HCl, pH 6.8, 20% glycerol, 4% SDS, 0.2% bromphenol blue, 5% β-mercaptoethanol). Total proteins were separated in a 4–15% SDS-PAGE gradient and transferred to nitrocellulose membrane (GE Healthcare Life Sciences). Membranes were blocked with 5% (m/v) skimmed milk powder at room temperature for 1 hr and then incubated with primary antibodies at 37°C for 1 hr. After washing three times, membranes were incubated with secondary antibodies at 37°C for 1 hr. Antibodies against RFP (1:3000), P (1:3000), GST (1:5000), GFP (1:5000; MBL, 598), Flag (1:5, 000; Sigma, F1804) were used for protein detection. Goat anti-rabbit IgG (EASYBIO, BE0101) and goat anti-mouse IgG horseradish peroxidase conjugate (Bio-Rad, 170–6516) were used as secondary antibodies. After addition of NcmECL Ultra stabilized peroxide reagent (NCM Biotech, P10300B), chemiluminescence of membranes was detected with a Biomolecular Imager (Azure biosystems, Inc).
4
Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plant samples were ground in liquid nitrogen, and total proteins were isolated by extraction buffer [0.2 M NaCl, 5 mM MgCl2, 5 mM DTT, 20 mM tris-HCl (pH 7.5), 0.03% Tween 20 (Ameresco), and 0.5 tablets of protease inhibitor (Roche)]. The supernatant was collected by centrifuging at 12,000 rpm for 15 min. Total proteins were examined by Western blot analysis using α-tubulin (1:5000; EASYBIO, BE0031) as a loading control.
Proteins in the study were also probed with α-RH12 (1:2000; GenScript, GTGRGAPPNPDYHQC), α-SE (1:2000; Agrisera, AS09532A), α-HYL1 (1:2000; Agrisera, AS06136), α-GFP (1:2000; EASYBIO, BE2001), α-FLAG (1:2000; Sigma-Aldrich, F7425), α-HA (1:2000; EASYBIO, BE7002), α-TuMV-VPg (1:200; GenScript, KGKSKGRTRGIGHKC), α-TuMV-CP (1:5000; GenScript, AGETLDAGLTDEQKC), α-TuMV-HC-Pro (1:2000), α-CMV-CP (1:3000), and α-MEMB12 (1:2000) (23 (link)). Secondary antibodies were goat anti-rabbit IgG (1:5000; EASYBIO, BE0101) and goat anti-mouse IgG (1:5000; EASYBIO, BE0102).
Proteins in the study were also probed with α-RH12 (1:2000; GenScript, GTGRGAPPNPDYHQC), α-SE (1:2000; Agrisera, AS09532A), α-HYL1 (1:2000; Agrisera, AS06136), α-GFP (1:2000; EASYBIO, BE2001), α-FLAG (1:2000; Sigma-Aldrich, F7425), α-HA (1:2000; EASYBIO, BE7002), α-TuMV-VPg (1:200; GenScript, KGKSKGRTRGIGHKC), α-TuMV-CP (1:5000; GenScript, AGETLDAGLTDEQKC), α-TuMV-HC-Pro (1:2000), α-CMV-CP (1:3000), and α-MEMB12 (1:2000) (23 (link)). Secondary antibodies were goat anti-rabbit IgG (1:5000; EASYBIO, BE0101) and goat anti-mouse IgG (1:5000; EASYBIO, BE0102).
5
Protein Analysis of Renal Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Renal tissues or cells were homogenized in RIPA lysis buffer (Mei5bio) containing 4% protease inhibitor cocktail (Roche, Basel, Switzerland) and 1% phosphatase inhibitor (Applygen, Beijing, China). Total protein concentration was detected by BCA kit (Pierce, Rockford, IL, USA) after ultrasonic cracking. Identical amounts of protein samples were electrophoresed on polyacrylamide gels and electrotransferred to polyvinylidene difluoride membranes, and then the membranes were incubated with antibodies against b-actin (Santa Cruz), COX-2, IL-6, TLR4 (ABclonal, Wuhan, China), iNOS, MyD88 (Abcam, Cambridge, UK), caspase-3, cleaved caspase-3, caspase-8, cleaved caspase-8, NF-kB, p-NF-kB (Immunoway, Plano, TX, USA), at 4 °C overnight. Then, goat anti-rabbit IgG or goat anti-mouse IgG (EASYBIO, Beijing, China) were added and the blots were developed with ECL plus kit (Biodragon, Beijing, China) and were visualized with a chemiluminescence detection system (Syngene, GeneGnome XRQ, Cambridge, UK). Quantitation was performed by scanning and analyzing the intensity of the hybridization bands and the data were analyzed with Image J software.
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!