Slice cultures were fixed with 4% paraformaldehyde overnight at 4 °C and incubated in blocking buffer (10% normal goat serum in 0.1% Triton-X in PBS) for 1 h. Cultures were left in primary antibody (1:200 anti-tau, abcam; cat. #ab75714), 1:500 anti-MAP2 (BD Pharmingen, San Diego, CA, USA; cat. #556320), 1:200 anti-DRP1 EPR19274 (abcam; cat. #ab184247), 1:200 anti-NCX1 EPR12739 (abcam; cat. #ab177952), and 1:200 anti-TOMM20 (abcam; cat. #ab56783) overnight at 4 °C, washed 3× with PBS, and incubated with respective secondary antibody conjugated to Alexa Fluor 488 or 647 (1:500; ThermoFisherScientific). Fluorescence micrographs were taken under a ×20 objective using the EVOS FL Microscope (ThermoFisherScientific). The following filter cubes were used for this study: EVOS™ Light Cube, GFP cat. AMEP4651; EVOS™ Light Cube, RFP cat. AMEP4652; EVOS™ Light Cube, Cy™5 cat. AMEP4656. The Look Up Table (LUT) applied to display CellROX Green fluorescence intensity is linear and found in ImageJ (16-color LUT). Colocalization of TOMM20 and Drp1 immunostaining was determined using the Coloc 2 plugin (https://imagej.net/Coloc_2 ) in ImageJ.
Anti drp1 epr19274
Manufactured by Abcam
Anti-DRP1 EPR19274 is a recombinant rabbit monoclonal antibody that recognizes the DRP1 (dynamin-related protein 1) protein. DRP1 is a GTPase that plays a key role in the regulation of mitochondrial fission.
Automatically generated - may contain errors
Lab products found in correlation
Anti tomm20, by Abcam (1 mentions)
Evos fl microscope, by Thermo Fisher Scientific (1 mentions)
Anti tau, by Abcam (1 mentions)
Alexa fluor 488 or 647, by Thermo Fisher Scientific (1 mentions)
Anti map2, by BD (1 mentions)
Las4000 imager, by GE Healthcare (1 mentions)
Immobilon p polyvinylidene difluoride, by Merck Group (1 mentions)
Anti tubulin, by Merck Group (1 mentions)
Mouse anti v5, by Thermo Fisher Scientific (1 mentions)
Anti gfp, by Thermo Fisher Scientific (1 mentions)
Ecl prime, by GE Healthcare (1 mentions)
Anti β actin, by Merck Group (1 mentions)
2 protocols using anti drp1 epr19274
1
Immunostaining of Neuronal Markers
2
Western Blot Analysis of Mitochondrial Proteins
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WB were performed by standard procedures using polyvinylidene fluoride membranes (Immobilon-P polyvinylidene difluoride, Millipore), ECL Prime (GE Healthcare) and analyzed using a LAS4000 Imager (GE Healthcare). Primary antibodies were used as follows: anti-SAMD4A HPA043061 (Sigma) 1:100; anti-SAMD4B HPA059385 (Sigma) 1:100, mouse anti-V5 (Invitrogen) 1:5,000, anti-GFP (Invitrogen) 1:2,000; antitubulin (DSHB) 1:10,000, anti-β-actin (Sigma-Aldrich) 1:10,000, anti-DRP1 EPR19274 (abcam) 1:1000, anti-OPA1 EPR11057(B) (abcam) 1:1000, anti-Mitofusin 2 6A8 (abcam) 1:1000, anti-TOM2O sc-17764 1:100. HRP-conjugated secondary antibodies were used 1:10,000 or 1:100,000. Signal intensity was assessed with the ImageJ software.
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