- PA5-29036 - Invitrogen Antibodies GPHN antibody

Gregor KM, Becker SC, Hellhammer F, Baumgärtner W, Puff C
Biology 2022 Jan 1;11(1)

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Gephyrin using 30 µg of A) 293T (B) A431 (C) HeLa and D) HepG2 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a Gephyrin polyclonal antibody (Product # PA5-29036) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Gephyrin using 30 µg of PC-12 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a Gephyrin polyclonal antibody (Product # PA5-29036) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of Gephyrin was performed by separating 50 µg of Various tissue extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a Gephyrin Polyclonal Antibody (Product # PA5-29036) at a dilution of 1:5000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Gephyrin Polyclonal Antibody (Product # PA5-29036). Sample (30 µg of whole cell lysate). Lane A: NIH-3T3. Lane B: JC . 7.5% SDS PAGE. Gephyrin Polyclonal Antibody (Product # PA5-29036) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of Gephyrin was performed by separating 30 µg of various whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a Gephyrin Polyclonal Antibody (Product # PA5-29036) at a dilution of 1:3000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Gephyrin Polyclonal Antibody (Product # PA5-29036). Human tissue extract (30 µg) was separated by 7.5% SDS-PAGE, and the membrane was blotted with Gephyrin Polyclonal Antibody (Product # PA5-29036) diluted at 1:2,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Gephyrin Polyclonal Antibody (Product # PA5-29036). Various tissue extracts (50 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with Gephyrin Polyclonal Antibody (Product # PA5-29036) diluted at 1:5,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of Gephyrin was achieved by transfecting HeLa with Gephyrin specific siRNAs (Silencer® select Product # s20000, s20001). Western blot analysis (Fig. a) was performed using whole cell extracts from the Gephyrin knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Gephyrin Polyclonal Antibody (Product # PA5-29036, 1:3000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to Gephryrin. An additional band at 70 kDa was also observed.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on 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) and tissue extracts of Mouse Brain (Lane 6) and Rat Brain (Lane 7). The blot was probed with Anti-Gephyrin Polyclonal Antibody (Product # PA5-29036, 1:3000 dilution) and detected by chemiluminescence using Goat anti Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 95 kDa band corresponding to Gephyrin was observed across all cell lines and tissue extracts tested. An additional band at 70 kDa was also observed in all cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of Gephyrin was performed in DIV10 rat E18 primary hippocampal neurons fixed in 4% paraformaldehyde at RT for 15 min. Green: Gephyrin Polyclonal Antibody (Product # PA5-29036) diluted at 1:500. Red: beta Tubulin 3/ Tuj1, stained by beta Tubulin 3/ Tuj1 antibody. Blue: Fluoroshield with DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of Gephyrin was performed using 70% confluent log phase Neuro-2a cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Gephyrin Rabbit Polyclonal Antibody (Product # PA5-29036) at 5 microgram/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 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). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry (Frozen) analysis of Gephyrin was performed in frozen sectioned E13.5 Rat brain tissue using Gephyrin Polyclonal Antibody (Product # PA5-29036) at a dilution of 1:250 (Green). Red: beta Tubulin 3/ TUJ1, a mature neuron marker, stained by beta Tubulin 3/ TUJ1 antibody diluted at 1:500. Blue: Fluoroshield with DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of paraffin-embedded mouse brain, using Gephyrin (Product # PA5-29036) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunoprecipitation of Gephyrin was performed in A431 whole cell extracts using 5 µg of Gephyrin Polyclonal Antibody (Product # PA5-29036). Samples were transferred to a membrane and probed with Gephyrin Polyclonal Antibody as a primary antibody and an HRP-conjugated anti-Rabbit IgG was used as a secondary antibody.

PA5-29036