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LearnMA5-17609
antibody from Invitrogen Antibodies
Targeting: SNAP25
bA416N4.2, dJ1068F16.2, RIC-4, RIC4, SEC9, SNAP, SNAP-25
Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Other assay [1]
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- Product number
- MA5-17609 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SNAP25 Monoclonal Antibody (SP12)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- MA5-17609 targets SNAP-25 in ELISA, FACS, and WB applications and shows reactivity with Human samples.
- Antibody clone number
- SP12
- Concentration
- 1 mg/mL
Submitted references Structural Basis for Substrate Recognition by the Ankyrin Repeat Domain of Human DHHC17 Palmitoyltransferase.
Verardi R, Kim JS, Ghirlando R, Banerjee A
Structure (London, England : 1993) 2017 Sep 5;25(9):1337-1347.e6
Structure (London, England : 1993) 2017 Sep 5;25(9):1337-1347.e6
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-SNAP25 Monoclonal Antibody (SP12) (Product # MA5-17609) and a 25 kDa band corresponding to Synaptosomal-associated protein 25 was observed across the cell lines tested. Whole cell 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) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:10,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Synaptosomal-associated protein 25 was performed using 70% confluent log phase PC-12 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with SNAP25 Monoclonal Antibody (SP12) (Product # MA5-17609) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing enhanced cytoplasmic and membrane localization of SNAP-25 in PC-12 cells differentiated to neuronal phenotype with NGF (200 ng/mL 5 days). Panel e represents the undifferentiated PC-12 cells. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
