Immunostaining of myofiber preparations was performed as previously described [37 (link)]. Briefly, myofiber preparations were fixed with 4% PFA, for 20 min, permeabilized with PBST (0.3% TritonX in PBS), blocked for 1 hour with PBSTB (5% BSA in PBT) and incubated overnight at 4°C with primary antibodies. The following primary antibodies were used: mouse anti-DHPR (1:200; DHPRa1A; Abcam), mouse anti-α-Actinin (1:100; Sigma), mouse anti-RyR1 (1:100; 34C; DSHB), rabbit anti-Junctin (1:350; gift from Dulhunty lab), mouse anti-PI(3)P (1:100; Echelon Biosciences Inc.), mouse anti-PI(3,4)P2 (1:100; Echelon Biosciences Inc.), mouse anti-PI(4,5)P2 (1:100; Echelon Biosciences Inc.). Alexa Fluor-conjugated secondary antibodies were used at 1:1000 (Invitrogen). Rhodamine phalloidin (Phalloidin 555) was used to visualize filamentous actin (1:300, Molecular Probes). Preparations were mounted with ProLong Gold with DAPI (Invitrogen). Images were acquired with a Nikon Eclipse Ti laser scanning confocal using NIS Elements software (Nikon Corporation, Tokyo, Japan) and only adjusted for brightness and contrast using Adobe Photoshop.
Dhpra1a
Manufactured by Abcam
DHPRa1A is a protein that is a subunit of the dihydropyridine receptor, a voltage-dependent calcium channel found in the sarcoplasmic reticulum of skeletal muscle cells. It plays a role in the regulation of calcium release and muscle contraction.
Automatically generated - may contain errors
Lab products found in correlation
Rhodamine phalloidin phalloidin 555, by Thermo Fisher Scientific (2 mentions)
Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (2 mentions)
Prolong gold with dapi, by Thermo Fisher Scientific (2 mentions)
Mouse anti pi 3 p, by Echelon Biosciences (2 mentions)
Mouse anti pi 3 4 p2, by Echelon Biosciences (2 mentions)
Mouse anti pi 4 5 p2, by Echelon Biosciences (2 mentions)
Mouse anti α actinin, by Merck Group (1 mentions)
Nis elements software, by Nikon (1 mentions)
Mouse anti actinin, by Merck Group (1 mentions)
Eclipse ti, by Nikon (1 mentions)
2 protocols using dhpra1a
1
Immunostaining of Myofiber Preparations
2
Immunostaining of Myofiber Preparations
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Immunostaining of myofiber preparations was performed as previously described (Horstick et al., 2013) . Briefly, myofiber preparations were fixed with 4% PFA, for 20 min, permeabilized with PBST (0.3% TritonX in PBS), blocked for 1 hour with PBSTB (5% BSA in PBT) and incubated overnight at 4 o C with primary antibodies. The following primary antibodies were used: mouse anti-DHPR (1:200; DHPRa1A; Abcam), mouse anti--Actinin (1:100; Sigma), mouse anti-RyR1 (1:100; 34C; DSHB), rabbit anti-Junctin (1:350; gift from Dulhunty lab), mouse anti-PI(3)P (1:100; Echelon Biosciences Inc.), mouse anti-PI(3,4)P2 (1:100; Echelon Biosciences Inc.), mouse anti-PI(4,5)P2 (1:100; Echelon Biosciences Inc.). Alexa Fluor-conjugated secondary antibodies were used at 1:1000 (Invitrogen). Rhodamine phalloidin (Phalloidin 555) was used to visualize filamentous actin (1:300, Molecular Probes). Preparations were mounted with ProLong Gold with DAPI (Invitrogen). Images were acquired with a Nikon Eclipse Ti laser scanning confocal using NIS Elements software (company, location) and only adjusted for brightness and contrast using Adobe Photoshop.
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