Tris, sodium dodecyl sulfate (SDS), 30% acrylamide and protein lysis buffer (#R0010) were obtained from Solarbio Science & Technology Company (Beijing, China). TRzol was purchased from Ambion company (Texas, United States). Anti-CD11b (#ab1211, #ab133357), anti-IL-1β (#ab9787), anti-CD40 (#ab13545), anti-CD68 (#ab125212), and anti-α7nAChR (#ab10096) were purchased from Abcam (Cambridge, United Kingdom). Anti-IKK (#2682s), anti-phospho-IKK(#2697s), anti-P65-NF-κB (#242s), anti-phospho-P65--NF-κB (#3033s), anti-IκB (#4814s) and anti-phospho-IκB (#9246s) were purchased from Cell Signaling Technology (Massachusetts, United States). Anti-β-tubulin (#MA511732), AlexaFluor488 mouse secondary antibody (#A11029), AlexaFluor488 rabbit secondary antibody (#A11034), enhanced chemiluminescence reagent (ECL) Western blotting detection reagents (#32106), Reverse Transcription Master Mix kit (Invitrogen, #4374966), SYBR® Select Master Mix (Life Technologies, #4472908), 0.25% trypsin (Gibco, #25200-056), DMEM medium (Gibco, #31800-014), fetal bovine serum (FBS) (Gibco, #10099-141) and penicillin streptomycin combination (Gibco, #15140-122) were purchased from Thermo Fisher Scientific (Massachusetts, United States). 2-, 3-, 5-triphenyltetrazolium chloride (TTC, #T8877-25G) was purchased from Sigma (United States).
Anti α7nachr
Manufactured by Abcam
Sourced in United States
Anti-α7nAChR is a laboratory research reagent used for the detection and study of the α7 nicotinic acetylcholine receptor (α7nAChR) in various biological samples. It functions as a specific antibody that binds to the α7nAChR protein, enabling its identification and quantification through various immunodetection techniques.
Automatically generated - may contain errors
Lab products found in correlation
Protein lysis buffer, by Solarbio (2 mentions)
Penicillin streptomycin combination, by Thermo Fisher Scientific (2 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
Trzol, by Thermo Fisher Scientific (2 mentions)
Anti phospho p65 nf κb 3033s, by Cell Signaling Technology (2 mentions)
Anti β tubulin, by Thermo Fisher Scientific (2 mentions)
Alexafluor488 mouse secondary antibody, by Thermo Fisher Scientific (2 mentions)
Alexafluor488 rabbit secondary antibody, by Thermo Fisher Scientific (2 mentions)
Reverse transcription master mix kit, by Thermo Fisher Scientific (2 mentions)
Anti phospho iκb, by Cell Signaling Technology (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dmem medium, by Thermo Fisher Scientific (2 mentions)
Anti iκb, by Cell Signaling Technology (1 mentions)
Anti il 1β, by Abcam (1 mentions)
Anti cd68, by Abcam (1 mentions)
Sybr select master mix, by Thermo Fisher Scientific (1 mentions)
Anti mpo, by Santa Cruz Biotechnology (1 mentions)
Cm3050, by Leica (1 mentions)
Anti iba1, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions)
6 protocols using anti α7nachr
1
Molecular Mechanisms of Microglial Activation
2
Neuroinflammation and Nicotinic Receptor Evaluation
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At 24 hours after surgery, anesthetized animals were transcardially perfused with PBS followed by 4% paraformaldehyde. The brains were removed, fixed with formalin (for 3 days), and dehydrated with 30% sucrose solution (for 3 days). Brain specimens were then flash frozen and cut into 10 μm slices using a cryostat (CM3050S, Leica Biosystems, Inc., IL, USA). All brain slices between 1 mm anterior and posterior from the center of the hemorrhage were washed with PBS and then incubated with blocking solution (10% normal goat serum in 0.1 M PBS). The slides were incubated with the following primary antibodies: anti-α7nAChR and anti-Iba1 (Abcam, MA, USA), as well as anti-MPO (Santa Cruz Biotechnology, TX, USA), and DAPI (Vector Laboratories, Inc., CA, USA). After washing the slides in PBS, the appropriate secondary antibodies (Santa Cruz Biotechnology, TX, USA) were applied for 2 hours at room temperature. Stained slices were evaluated using an OLYMPUS BX51 microscope.
3
Hippocampal Protein Expression Analysis
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Hippocampi were separated, homogenized with ice-cold RIPA buffer containing 1% PMSF, and centrifuged at 12,000 g for 10 min at 4°C. The supernatant was collected and the protein concentration was measured using a BCA kit (Beyotime, Shanghai, China). Protein samples (30 μg each) were isolated by 10% SDS-PAGE and transferred to PVDF membranes (Millipore, United States ). After blocking with 5% nonfat milk in Tris-buffered saline containing Tween 20 (TBST) at room temperature for 1 h, membranes were incubated with anti-α7nAChR (1:500, Abcam, Cat.10096, United States), anti-GLT-1 (1:1,000, Invitrogen, Cat.701,988, United States), and anti-SYP (1:1,000, Servicebio, Cat.11553, China) antibodies overnight at 4°C. The membranes were washed three times with TBST and were then subsequently incubated with HRP-conjugated secondary antibodies in TBST at room temperature for 1 h. The membrane was dyed with ECL reagent and developed using autoradiography film. Protein-band intensities were quantified using Quantity One software (Bio-Rad).
4
Cholinergic System Immunohistochemistry
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Prepared sections were reacted with anti-ACh (Chemicon), anti-VAChT (Abcam), anti-α7 nAChR (Abcam) and anti-AChE (Novus) antibodies. An immunohistochemical staining secondary antibodies kit (Maixin Biotechnology, Inc., Fuzhou China) was used according to the manufacturer’s instructions, and then colored with DAB (Maixin Biotechnology, Inc., Fuzhou China). Sections were counterstained with hematoxylin before they were covered. Immunohistochemical staining imaging was captured using an OLYMPUS BX61 (Tokyo, Japan) optical microscope.
5
Molecular Mechanisms of Neuroinflammation
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Tris, sodium dodecyl sulfate (SDS), 30% acrylamide and protein lysis buffer (#R0010) were obtained from Solarbio Science & Technology Company (Beijing, China). TRzol was purchsed from Ambion company (Texas, United States). Anti-CD11b (#ab1211), anti-IL-1β (#ab9787), anti-CD40 (#ab13545), anti-CD68 (#ab31630), and anti-α7nAChR (#ab10096) were purchased from Abcam (Cambridge, United Kingdom). Anti-IKK (#2682s), anti-phospho-IKK(#2697s), anti-P65-NF-κB (#242s), anti-phospho-P65--NF-κB (#3033s), anti-IκB(#4814s) and anti-phospho-IκB (#9246s) were purchased from Cell Signaling Technology (Massachusetts, United States). Anti-β-tubulin (#MA511732), AlexaFluor488 mouse secondary antibody (#A11029), AlexaFluor488 rabbit secondary antibody (#A11034), enhanced chemiluminescence reagent (ECL) Western blotting detection reagents (#32106), Reverse Transcription Master Mix kit (Invitrogen, #4374966), SYBR Ò Select Master Mix (Life Technologies, #4472908), 0.25% trypsin (Gibco, #25200-056), DMEM medium (Gibco, #31800-014), fetal bovine serum (FBS) (Gibco, #10099-141) and penicillin streptomycin combination (Gibco, #15140-122) were purchased from Thermo Fisher Scienti c (Massachusetts, United States). 2-, 3-, 5-triphenyltetrazolium chloride (TTC, #T8877-25G) was purchased from Sigma (United States).
6
Western Blot Analysis of Neurotransmitter Receptors
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Tissues were homogenized using lysis buffer (Beyotime, China), and supernatants were collected after centrifugation at 12,000 × g for 15 min at 4°C. After quantitative analysis of protein concentration, total proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to polyvinylidene fluoride membranes (Millipore, Billerica, Mass), blocked with 5% non-fat milk in Tris-buffered saline for 1 h at 37°C, and then incubated overnight at 4°C with primary antibodies. After incubation for 1 h at 37°C with secondary antibody (1:2,000; Beyotime), bands were seen using the enhanced chemilumine-scence kit (Beyotime) according to the manufacturer's protocol. The primary antibodies used were as follows: antineuregulin-1 (1:500, sc-28916, Santa Cruz Biotechnology Inc, Dallas, Tex), anti-γ-nAChR (1:500, sc-13998, Santa Cruz Biotechnology Inc, Dallas, Tex), anti-α7-nAChR (1:1,000, ab10096, Abcam, Ltd, Cambridge, Mass), anti-LC3II (1:500, 4108, Cell Signaling Technology, Danvers, Mass), anti-p62 (1:500, 5114, Cell Signaling Technology, Danvers, Mass), anti-GAPDH (1:1,000, AF1186, Beyotime Institute of Biotechnology, Jiangsu Province, China). All results were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) levels.
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