All substances were purchased from Sigma-Aldrich (Taufkirchen, Germany) unless stated otherwise. Papain was obtained from Roche (Mannheim, Germany). Chloromethyl-tetramethyl-rosamine (Mitotracker Orange) was purchased from Molecular Probes (Life Technologies, Carlsbad, CA, USA). For immunofluorescence staining, the following primary antibodies were used: mouse anti-glial fibrillary acidic protein (GFAP; 1:200; G-A-5 clone, Sigma), goat anti-calretinin (1:500, Swant, Marly, Switzerland), rabbit-anti-Aif1 (1:500, Wako Chemicals), mouse anti-glutamine synthetase (1:1000, Merck Millipore, Darmstadt, Germany), rabbit anti-TSPO (1:100, Abcam, Cambridge, UK), and rabbit anti-DBI (1:200, Sigma). As secondary antibodies, we used Cy5-conjugated donkey anti-goat, Cy3-conjugated donkey anti-rabbit, Cy2-conjugated donkey anti-mouse, Cy3-conjugated goat anti-rabbit, and Cy2-conjugated goat anti-mouse. All secondary antibodies were obtained from Dianova (Hamburg, Germany) and applied at 1:200 dilution.
Rabbit anti tspo
Manufactured by Abcam
Sourced in Germany, United States, United Kingdom
Rabbit anti-TSPO is a primary antibody that recognizes the translocator protein (TSPO), also known as the peripheral benzodiazepine receptor. TSPO is a widely expressed protein involved in various cellular processes, including cholesterol transport and steroidogenesis. This antibody can be used for the detection and study of TSPO in various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
Bca protein assay reagent, by Thermo Fisher Scientific (2 mentions)
Las 4000, by Fujifilm (2 mentions)
Complete protease inhibitor, by Roche (2 mentions)
Papain, by Roche (1 mentions)
Goat anti calretinin, by Swant (1 mentions)
Mouse anti glutamine synthetase, by Merck Group (1 mentions)
Gel pro analyzer software version 3, by Media Cybernetics (1 mentions)
Protease inhibitor and phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Ripa lysis buffer, by Applygen (1 mentions)
Western lightning plus enhanced chemoluminescence system, by PerkinElmer (1 mentions)
Enhanced chemoluminescence system, by PerkinElmer (1 mentions)
Cresyl violet, by Merck Group (1 mentions)
Mouse anti neun, by Merck Group (1 mentions)
Rabbit anti iba1, by Fujifilm (1 mentions)
Tmr red, by Roche (1 mentions)
5 protocols using rabbit anti tspo
1
Immunofluorescence Staining of Glial Markers
2
Quantitative Western Blot Analysis of TSPO
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The punched hippocampal tissues (3 mm in diameter around the injection sites on both sides) were extracted by RIPA lysis buffer (Applygen, China) plus protease inhibitor and phosphatase inhibitor cocktail (Thermo Pierce, Rockford, IL, USA). Equal amounts (50 μg) of protein were resolved by electrophoresis, transferred onto polyvinylidene difluoride (PVDF) membranes and blocked with 5% skim milk solution. The membranes were incubated with rabbit anti-TSPO (at a dilution of 1:1000; Abcam, Cambridge, MA), or β-actin (1:3000; Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 4°C overnight. After washing and incubating with secondary antibodies (1:3000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), the specific bands were detected and quantified using Gel-Pro Analyzer software, Version 3.1 (Media Cybernetics, Bethesda, MD, USA).
3
Western Blotting of TSPO in Mouse Brains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Adult mice brains were dissected and reduced in powder at −80 °C, immediately dissolved in PBS with 2% SDS, and 1× EDTA-free Complete Protease Inhibitor (Roche, Basel, Switzerland). Lysates were sonicated twice at 10 Hz (Vibra cell VCX130) and centrifuged for 30 min at 16,000 rcf at 4 °C. Supernatants were boiled in 5× Laemmli loading buffer. Protein content was measured using the BCA protein assay reagent (Thermo scientific, Waltham, MA, USA). Equal amounts of proteins (20 µg) were separated by denaturing electrophoresis in NuPAGE 4–12% Bis-Tris acetate gradient gel (Invitrogen, Carlsbad, CA, USA) and electrotransfered to nitrocellulose membranes. Membranes were analyzed using the following primary antibodies: rabbit anti-TSPO (Abcam, Cambridge, UK) (1:10,000); horseradish-peroxidase-conjugated (HRP) anti-GAPDH (1:10,000); Secondary antibody used was HRP-conjugated anti-rabbit antibodies (Amersham) (dilution 1:2000). HRP activity was visualized by enhanced chemiluminescence (ECL) using Western Lightning plus enhanced chemoluminescence system (Perkin Elmer, Waltham, MA, USA). Chemoluminescence imaging was performed on a LAS4000 (Fujifilm, Tokyo, Japan). GAPDH expression was used as a loading reference.
4
Brain Region Protein Extraction and Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After imaging, mice were sacrificed and their brain regions (cortex and hippocampus) dissected. Samples were reduced in powder at −80 °C and immediately dissolved in PBS with 2% SDS and 1× EDTA-free Complete Protease Inhibitor (Roche®). The lysates were sonicated twice at 10 Hz (Vibra cell VCX130) and centrifuged for 30 min at 16,000× g at 4 °C. Supernatants were boiled in Laemmli loading buffer. Protein content was measured using the BCA protein assay reagent (Thermo Fisher®, Les Ulis, France). Equal amounts of proteins (20 µg) were separated by denaturing electrophoresis in NuPAGE® 4–12% Bis-Tris acetate gradient gel (Thermo Fisher®) and electro-transferred to nitrocellulose membranes. The membranes were analyzed using the following primary antibodies: rabbit anti-TSPO (Abcam) (1:10,000); horseradish-peroxidase-conjugated (HRP) anti-GAPDH (1:10,000); and the secondary antibodies, HRP-conjugated anti-rabbit (GE Healthcare) (dilution 1:2000). HRP activity was visualized by enhanced chemo-luminescence (ECL) using Western Lightning plus an enhanced chemoluminescence system (Perkin Elmer®, Waltham, MA, USA). Chemoluminescence imaging was performed on a LAS4000 (Fujifilm®,Tokyo Japan). GAPDH expression was used as a loading reference.
5
Evaluating Thalamic Damage After dMCAO
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Eight rats per group were perfused 7 or 14 days after dMCAO. The brains were removed from the skulls and postfixed. Coronal sections (10-μm thick) were cut and stored at -80°C. Every 12th coronal section from bregma +4.7 to -5.2 mm was selected for Nissl staining using 0.3% cresyl violet (Sigma) to evaluate infarct volumes. Every sixth coronal section from bregma -1.8 to -3.6 mm was selected to assess secondary damage in the thalamus and hippocampus, and thalamic TSPO expression and Aβ deposition using Nissl and immunohistochemical staining. We used rabbit anti-TSPO (1:100; Abcam), mouse anti-NeuN (neuronal nuclei, 1:200; Millipore), mouse anti-GFAP (glial fibrillary acidic protein, 1:500; CST), rabbit anti-Iba-1 (ionized calcium-binding adaptor molecule-1, 1:400; Wako), or rabbit anti-Aβ 3-16 (1:300; Covance). Primary antibody incubation was followed by incubation in secondary Alexa Fluor 488/555-conjugated anti-mouse (1:500; CST) or anti-rabbit (1:500; CST) antibodies. We performed terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) using an in situ cell death detection kit, TMR red (Roche) according to the manufacturer's suggestions to examine apoptosis.
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!