Tissue was dissociated with FACS lysis buffer (final concentration: 0.32M Sucrose, 10mM Tris − HCl pH8.0, 5mM CaCl2, 3mM Mg(acetate)2, 0.1mM EDTA, 1mM DTT, 0.3% Triton-X −100 and 100× PIC) into single cell suspension, then fixed with 1% formaldehyde for 5 mins, and stop by 0.125M Glycine. Then, cells were washed twice with cold 1xPBS to remove excessed formaldehyde and glycine. After incubating with blocking buffer (Final concentration: 10% normal goat serum, 5% BSA, 0.1% Triton-X-100 and 1XPIC) for half-hour, cells were double-labeled with Arc antibody (Bioss) in 1:20000 dilution per million cell and NeuN antibody (Abcam) in 1:20000 per million cell, together with DAPI (Thermofisher) in 1:2000. PBPT buffer was used for washing (twice each time) and resuspend into 500ul 1x PBS for FACS sorting. FACS was performed on a BD FACSArial (BD Science), and cells were sorted into lysis buffer from Arcturus PicoPure RNA isolation kit (Applied Biosystems). Then, RNA was isolated from sorted cells by Arcturus PicoPure RNA isolation kit (Applied Biosystems) following the manufacturers protocol. 5ng of total RNA per sample and SMRTer Stranded Total RNA-seq Kit v2 Pico Input Mammalian Components kit (Clontech) was used for RNA-seq library preparation following the manufacturers protocol. Sequencing was conducted at GENEWIZ (Suzhou, China) with 150pb Pair-End reads.
Arc antibody
Manufactured by Bioss Antibodies
The Arc antibody is a laboratory reagent used to detect and study the Arc protein in biological samples. The Arc protein is involved in various cellular processes, including synaptic plasticity and memory formation. The Arc antibody can be used in techniques such as Western blotting, immunohistochemistry, and immunoprecipitation to identify and quantify the Arc protein.
Automatically generated - may contain errors
2 protocols using arc antibody
1
Single-cell RNA-seq of FACS-sorted Arc+ Neurons
2
Single-cell RNA-seq of FACS-sorted Arc+ Neurons
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Tissue was dissociated with FACS lysis buffer (final concentration: 0.32M Sucrose, 10mM Tris − HCl pH8.0, 5mM CaCl2, 3mM Mg(acetate)2, 0.1mM EDTA, 1mM DTT, 0.3% Triton-X −100 and 100× PIC) into single cell suspension, then fixed with 1% formaldehyde for 5 mins, and stop by 0.125M Glycine. Then, cells were washed twice with cold 1xPBS to remove excessed formaldehyde and glycine. After incubating with blocking buffer (Final concentration: 10% normal goat serum, 5% BSA, 0.1% Triton-X-100 and 1XPIC) for half-hour, cells were double-labeled with Arc antibody (Bioss) in 1:20000 dilution per million cell and NeuN antibody (Abcam) in 1:20000 per million cell, together with DAPI (Thermofisher) in 1:2000. PBPT buffer was used for washing (twice each time) and resuspend into 500ul 1x PBS for FACS sorting. FACS was performed on a BD FACSArial (BD Science), and cells were sorted into lysis buffer from Arcturus PicoPure RNA isolation kit (Applied Biosystems). Then, RNA was isolated from sorted cells by Arcturus PicoPure RNA isolation kit (Applied Biosystems) following the manufacturers protocol. 5ng of total RNA per sample and SMRTer Stranded Total RNA-seq Kit v2 Pico Input Mammalian Components kit (Clontech) was used for RNA-seq library preparation following the manufacturers protocol. Sequencing was conducted at GENEWIZ (Suzhou, China) with 150pb Pair-End reads.
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