Pyrrolidine dithiocarbamate (PDTC), minocycline and fluorocitrate were obtained from Sigma (St. Louis, MO, USA). The normal goat IgG, anti-CCL5 neutralizing antibody and recombinant rat CCL5 were purchased from R&D Systems (Minneapolis, MN, USA). Anti-rat CCL5, rabbit anti-rat NF-κB p65 and mouse anti-rat β-actin were obtained from Santa Cruz (Santa Cruz, CA, USA). Fluorescein isothiocyanate (FITC)-conjugated IgG and tetraethyl rhodamine isothiocyanate (Jackson Immunolab, West Grove, PA, USA), glial fibrillary acidic protein (GFAP, Millipore, Bedford, MA, USA), ionized calcium–binding adapter molecule 1 (Iba-1, Abcam), and neuronal specific nuclear protein (NeuN, neuronal marker, NOVUS) were purchased. The dosages of intrathecal drugs and peptides were chosen according to former studies [17 (link), 29 (link)] and our preliminary tests.
Mouse anti rat β actin
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Mouse anti-rat β-actin is a monoclonal antibody that specifically binds to the β-actin protein in rat samples. It is a commonly used tool in molecular and cellular biology research.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (2 mentions)
Fluorocitrate, by Merck Group (1 mentions)
Normal goat igg, by R&D Systems (1 mentions)
Iba 1, by Abcam (1 mentions)
Glial fibrillary acidic protein (gfap), by Merck Group (1 mentions)
Goat anti rat il 6, by R&D Systems (1 mentions)
Quantity one software, by Bio-Rad (1 mentions)
Hmgb1, by Santa Cruz Biotechnology (1 mentions)
Complete protease inhibitor mixture tablet, by Roche (1 mentions)
Enhanced chemiluminescence, by Vilber (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Rockland Immunochemicals (1 mentions)
Fusion fx, by Vilber (1 mentions)
4 protocols using mouse anti rat β actin
1
Intrathecal Drug Delivery for Neuroinflammation
2
Protein Expression Analysis in Spinal Cord
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The L2 segment spinal cord was lysed with the lysis buffer. After homogenizing, standing, and centrifugation, the protein concentration was determined. The protein samples underwent analysis with 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and transferred onto the SDS-PAGE membrane. After blocking with 10% dried skimmed milk powder at 4°C overnight, the membrane was incubated for 1 h with the primary antibodies, including mouse anti-rat CNTF primary antibody (1: 500) (Chemicon International Inc., Temecula, CA, USA), goat anti-rat IL-6 (1: 200) (R&D Systems, Minneapolis, MN, USA), goat anti-rat IL-10 (1: 2000 dilution; R&D Systems), and mouse anti-rat β-actin (1: 2000) (Santa Cruz Biotechnology Inc., Dallas, TX, USA). The membrane was incubated with secondary antibody conjugated with horseradish peroxidase (HRP), goat anti-mouse IgG (1: 5,000) (Santa Cruz Biotechnology Inc., Dallas, TX, USA) at room temperature for 70 min. Color development was performed with the enhanced chemiluminescence (ECL) kit, and protein band images were obtained and analyzed with Quantity One software (Bio-Rad, Hercules, CA, USA). β-actin was used as the internal reference.
3
Frozen Testicular Tissue Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Frozen testicular tissue was prepared with lysis buffer (150 mM NaCl, 20 mM Tris HCl [pH 7.4], 0.1% sodium dodecyl sulfate [SDS], 1.0% Nonidet P-40, 0.5% sodium-deoxycholate, 0.2 mM phenylmethylsulfonyl fluoride, protease inhibitor cocktails, and phosphates inhibitors). Equal amounts of proteins were separated by 10% SDS-polyacrylamide gel electrophoresis (PAGE), and electrotransferred to polyvinylidene fluoride (PVDF) membranes (Millipore, USA). The proteins were then blocked with 5% BSA for 2 h at room temperature. The membranes were incubated with appropriately diluted rabbit anti-rat primary antibodies for the detection of HO-1 (Abcam, UK), MFN-2 (Boshide, Wuhan, China), HMGB-1 (Santa Cruz Biotechnology Co., USA), and mouse anti-rat β-actin (Santa Cruz Biotechnology Co., USA) overnight at 4°C. The membranes were then incubated with the relevant secondary antibodies (HO-1, MFN-2, HMGB-1: anti-rabbit IgG, β-actin: anti-mouse IgG; all purchased from Boshide, Wuhan, China) for 2 h at 20°C. The proteins were detected with an enhanced chemiluminescence (ECL) detection system. β-Actin served as the internal control. The analysis was repeated thrice.
4
Granulosa-Luteal Cell Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Granulosa-luteal cells (GLCs) were isolated from the follicular fluid of 68 women participating in the study. BMP-6 protein and PPARγ protein were measured using Western blotting. GLCs were Lysate with 50 mM Tris-HCl (pH 7.5), 150 mM NaCl, 0.5% sodium deoxycholic acid,1% Nonidet P-40, and complete protease inhibitor mixture tablets (Roche Applied Science, Penzberg, Germany). From each sample, 10 ng of protein were separated by 10% SDS-PAGE and electro-transferred to a PVDF membrane (EMD Millipore, USA). Membranes were blocked with 5% milk for 2h at 25 C and incubated 12h at 4 C using the following primary antibodies: Rabbit anti-rat BMP-6 (1:1,000; cat No. ab155963; Abcam, UK), and Anti-PPARγ (Antibody (B-5): sc-271392, Santa Cruz Biotechnology, USA). The BMP-6 protein level and PPARγ protein level were normalized based on the level of β-actin protein. The primary antibody for β-actin was mouse anti-rat β-actin (1:1,000; cat no.sc-130656; Santa Cruz Biotechnology, USA). Membranes were incubated with horseradish peroxidase-conjugated secondary antibody (1:10,000, cat no.611-1302; Rockland) for 2h at 25 C and then evaluated with Enhanced Chemi-Luminescence, and Fusion Fx (Vilber Lourmat, France).
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!