Anti-Synaptojanin 1 antibody [RP23040243]

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Synaptojanin 1 antibody [RP23040243] (AB308136)

Immunohistochemical analysis 4% PFA-fixed, 0.1% Triton-X 100-permeabilized mouse brain tissue labelling Synaptojainin 1 with ab308136 at 1/250 dilution, followed by a Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate at 1 : 2000. Nuclei (blue) were stained with DAPI Panel (a) represents staining with the matched isotype control. Panel (b) shows a representative section stained for Synaptojainin 1 (green). The images were captured at 20X magnification.

• WB

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Western blot - Anti-Synaptojanin 1 antibody [RP23040243] (AB308136)

Western blot analysis was performed on Membrane enriched extracts (30 µg lysate) of PC-12 (Lane 1), PC-12 treated with NGF and KCl (NGF at 200 nM, KCl at 50 nM for 7 days) (Lane 2) and tissue extracts of Mouse Brain (Lane 3). The blots were probed with Anti-Synaptojanin Recombinant Rabbit Polyclonal Antibody (ab308136, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (0.25 µg/mL, 1 : 4000 dilution). A 173 kDa band corresponding to Synaptojanin was observed specifically in NGF differentiated PC-12 and tissue tested. Known quantity of protein samples were electrophoresed using a 4-12% Bis-Tris gel, electrophoresis system and pre-stained protein standard. Resolved proteins were then transferred onto a nitrocellulose membrane. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed (ECL).

Lane 1:

Membrane enriched extracts of PC-12 cell lysate at 30 µg

Lane 2:

Membrane enriched extracts of PC-12 cell lysate treated with NGF and KCl (NGF at 200 nM, KCl at 50 nM for 7 days) at 30 µg

Lane 3:

Mouse Brain tissue lysates at 30 µg

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/4000 dilution

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