- PA5-29116 - Invitrogen Antibodies DLG3 antibody

Vila A, Shihabeddin E, Zhang Z, Santhanam A, Ribelayga CP, O'Brien J
Frontiers in cellular neuroscience 2021;15:667046

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of SAP102 was performed by separating 30 µg of Molt-4 lysates by 7.5% SDS PAGE. Proteins were transferred to a membrane and probed with a SAP102 Polyclonal Antibody (Product # PA5-29116) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using SAP102 Polyclonal Antibody (Product # PA5-29116). Sample (50 µg of whole cell lysate). Lane A: Mouse brain . 7.5% SDS PAGE. SAP102 Polyclonal Antibody (Product # PA5-29116) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of SAP102 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 SAP102 Polyclonal Antibody (Product # PA5-29116) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-SAP102 Polyclonal Antibody (Product # PA5-29116) and a 90 kDa band corresponding to Disks large homolog 3 was observed across the neuronal cell lines tested and only in Mouse and Rat Brain which are reported to be positive and not in any other tissues. Whole cell extracts (30 µg lysate) of SH-SY5Y (Lane 1), SK-N-SH (Lane 2), Neuro-2a (Lane 3), IMR-32 (Lane 4) and tissue extracts (30 µg lysate) of Mouse Brain (Lane 5), Rat Brain (Lane 6), Mouse Skeletal Muscle (Lane 7), Rat Skeletal Muscle (Lane 8), Mouse Heart (Lane 9) and Rat Heart (Lane 10) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) 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-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 Novex® ECL Reagent Kit (Product # WP20005). An uncharacterized band of ~35 kDa was also observed in Rat Brain, Mouse Skeletal Muscle and Rat Skeletal Muscle.

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 C2C12 (Lane 1), SK-N-AS (Lane 2), Neuro-2a (Lane 3), SH-SY5Y (Lane 4), IMR-32 (Lane 5) and A549 (Lane 6). The blot was probed with Anti-SAP102 Polyclonal Antibody (Product # PA5-29116, 1:2000 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 90 kDa band corresponding to SAP102 was observed across the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of SAP102 in methanol-fixed A431 cells using a SAP102 polyclonal antibody (Product # PA5-29116) at a 1:200 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of SAP102 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: SAP102 Polyclonal Antibody (Product # PA5-29116) diluted at 1:500. Red: alpha Tubulin, a cytoskeleton marker. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of SAP102 was performed in DIV9 rat E18 primary cortical neurons fixed in 4% paraformaldehyde at RT for 15 min. Green: SAP102 Polyclonal Antibody (Product # PA5-29116) diluted at 1:500. Red: beta Tubulin 3/ Tuj1, a neuron cell marker. Blue: Fluoroshield with DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of SAP102 was performed using 70% confluent log phase SH-SY5Y 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 SAP102 Rabbit Polyclonal Antibody(Product # PA5-29116) at 5 µg/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, 1:300). Panel d represents the merged image showing cytoplasmic and membrane 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)
Main image
Experimental details: Immunohistochemistry (Paraffin) analysis of SAP102 was performed in paraffin-embedded mouse brain tissue using SAP102 Polyclonal Antibody (Product # PA5-29116) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry (Paraffin) analysis of SAP102 was performed in paraffin-embedded mouse brain tissue using SAP102 Polyclonal Antibody (Product # PA5-29116) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry (Frozen) analysis of SAP102 was performed in frozen sectioned E13.5 Rat brain tissue using SAP102 Polyclonal Antibody (Product # PA5-29116) 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.

PA5-29116