PA5-51827

antibody from Invitrogen Antibodies

Targeting: GPHN KIAA1385

Antibody data

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Validation data

Product number: PA5-51827 - Provider product page
Provider: Invitrogen Antibodies
Product name: Gephyrin Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant full-length protein
Description: Immunogen sequence: ATIQEHGYPT INLGIVGDNP DDLLNALNEG ISRADVIITS GGVSMGEKDY LKQVLDIDLH AQIHFGRVFM KPGLPTTFAT LDIDGVRKII FALPGNPVSA VVTCNLFVVP ALRKMQGILD PRPTIIKARL SCDVKLDPRP EY Highest antigen sequence identity to the following orthologs: Mouse - 100%, Rat - 100%.
Reactivity: Human
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 0.05 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

GPHN protein structure - PA5-51827 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Gephyrin in Lane 1: Marker (kDa) 230, 110, 82, 49, 32, 26, 18; Lane 2: Human cell line RT-4; Lane 3: Human cell line U-251MG sp; Lane 4: Human plasma (IgG/HSA depleted); Lane 5: Human liver tissue.; Lane 6: Human tonsil tissue. tissue. Samples were probed using a Gephyrin Polyclonal Antibody (Product # PA5-51827).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Gephyrin was achieved by transfecting PC-3 with Gephyrin specific siRNAs (Silencer® select Product # s20000, s20001). Western blot analysis (Fig. a) was performed using Membrane enriched extracts from the Gephyrin knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Gephyrin Polyclonal Antibody (Product # PA5-51827, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Gephyrin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Gephyrin Polyclonal Antibody(Product # PA5-51827) and a 93, 150, 250kDa band corresponding to Gephyrin was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), IMR-32 (Lane 3), Neuro-2a (Lane 4), PC-3 (Lane 5), PC-12 (Lane 6), MCF7 (Lane 7), A549 (Lane 8), B16-F10 (Lane 9), Mouse Brain (Lane 9), Rat Brain (Lane 10) 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 (0.5 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).The 150 kDa and 250 kDa bands observed in PC-12 are reported to be SUMOylated form of GPHN. [doi: 10.1038/ncomms13365 (2016)].