Western Blot Customer Review Anti-VGF Antibody (STJ96661)
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Rating: Specificity: ANTIBODY CUSTOMER REVIEW Findings: “The STJ96661 Anti-VGF antibody had a good binding capacity, 15μg loading per well gave specific target bands in both SH-SY5Y cell lysate and rat hippocampus lysate. .” – Wanjia Yu, University of St Andrews Knowledge Dock Business Centre, University Way, London E16 2RD T: (+44) 0208 223 3081 F: (+44) 0207 681 2580 E: [email protected] w: www.stjohnslabs.com Protocol Gel Electrophoresis 10% polyacrylamide gel, constant voltage start with 80 V for 40 min, then transfer to 120V for 30min. Membrane Wash 1X TBST wash for 3 times, 15min per time. Transfer 0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 25 V for 90 min. Secondary Antibody Probing Secondary antibody was added to the solution of 1X TBST with 5% BSA with 1:10000 dilution ratio, for 1 hr at RT. Blocking 1X TBST with 5% BSA for 1 hr at room temperature (RT). Membrane Wash 1X TBST wash for 3 times, 15min per time. Primary Antibody Probing Primary antibody was added to the solution of 1X TBST with 5% BSA, 4˚C for overnight. Visualisation ECL visualization for 2 min Model Number STJ96661 Antibody Name Anti-VGF antibody Application Western Blot Species Rat Tissue - Wild type SH-SY5Y cells; - ShFRMD6 SH-SY5Y cells (FRMD6 gene knock down cells); - Rat hippocampus lysate Dilution 1:1000 Secondary Antibody HRP goat anti-rabbit Secondary Dilution 1:10000 68kDa (VGF peptide precusor) Lane 1 2 3 4 5 6 7 8 9 Figure 1. Lane1: rat hippocampus; Lane 2-3: Wild type SH-SY5Y cell and ShFRMD6 SH- SY5Y cell lysate (without BDNF induction); Lane 4-5: Wild type SH-SY5Y cell and ShFRMD6 SH-SY5Y cell lysate (with BDNF induction for 6h); Lane 6-7: Wild type SH-SY5Y cell and ShFRMD6 SH-SY5Y precipitated cell culture medium (without BDNF induction); Lane 8-9: Wild type SH-SY5Y cell and ShFRMD6 SH-SY5Y precipitated cell culture medium (with BDNF induction for 6h)
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Transcript of Western Blot Customer Review Anti-VGF Antibody (STJ96661)
Rating:
Specificity:
ANTIBODY CUSTOMER REVIEWFindings: “The STJ96661 Anti-VGF antibody had a
good binding capacity, 15μg loading per well gave specific target bands in both SH-SY5Y cell lysate and rat hippocampus lysate. .” – Wanjia Yu, University of St Andrews
Knowledge Dock Business Centre, University Way, London E16 2RD T: (+44) 0208 223 3081 F: (+44) 0207 681 2580 E: [email protected] w: www.stjohnslabs.com
Protocol
Gel Electrophoresis
10% polyacrylamide gel, constant voltage start with 80 V for 40 min, then transfer to 120V for 30min.
Membrane Wash
1X TBST wash for 3 times, 15min per time.
Transfer 0.45 nm PVDF membrane, 1 piece of gel (wet transfer), constant voltage 25 V for 90 min.
Secondary Antibody
Probing
Secondary antibody was added to the solution of 1X TBST with 5% BSA with 1:10000 dilution ratio, for 1 hr at RT.
Blocking 1X TBST with 5% BSA for 1 hr at room temperature (RT).
Membrane Wash
1X TBST wash for 3 times, 15min per time.
Primary Antibody
Probing
Primary antibody was added to the solution of 1X TBST with 5% BSA, 4˚C for overnight.
Visualisation ECL visualization for 2 min
Model Number STJ96661
Antibody Name Anti-VGF antibody
Application Western Blot
Species Rat
Tissue - Wild type SH-SY5Y cells;- ShFRMD6 SH-SY5Y cells (FRMD6 gene knock down cells); - Rat hippocampus lysate
Dilution 1:1000
Secondary Antibody HRP goat anti-rabbit
Secondary Dilution 1:10000
68kDa(VGF peptide precusor)
Lane 1 2 3 4 5 6 7 8 9
Figure 1. Lane1: rat hippocampus; Lane 2-3: Wild type SH-SY5Y cell and ShFRMD6 SH-SY5Y cell lysate (without BDNF induction); Lane 4-5: Wild type SH-SY5Y cell and ShFRMD6 SH-SY5Y cell lysate (with BDNF induction for 6h); Lane 6-7: Wild type SH-SY5Y cell and ShFRMD6 SH-SY5Y precipitated cell culture medium (without BDNF induction); Lane 8-9: Wild type SH-SY5Y cell and ShFRMD6 SH-SY5Y precipitated cell culture medium (with BDNF induction for 6h)
