- PA5-21295 - Invitrogen Antibodies EPS8 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of EPS8 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 EPS8 Polyclonal Antibody (Product # PA5-21295) at a dilution of 1:1000. 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: Western Blot using EPS8 Polyclonal Antibody (Product # PA5-21295). Sample (30 µg of whole cell lysate). Lane A: H1299. Lane B: Hela . 7.5% SDS PAGE. EPS8 antibody . EPS8 Polyclonal Antibody (Product # PA5-21295) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-EPS8 Polyclonal Antibody (Product # PA5-21295) and 97 kDa band corresponding to EPS8 was increased upon LPS treatment in RAW 264.7 and was also expressed in COLO 205 when compared to HT-29. Membrane enriched extracts (30 µg lysate) of RAW 264.7 (Lane 1), RAW 264.7 treated with LPS (100 ng/mL, 24hr) (Lane 2), COLO 205 (Lane 3) and HT-29 (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). 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 Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 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)
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Experimental details: Immunofluorescence analysis of EPS8 was performed using 70% confluent log phase RAW 264.7 cells treated with LPS (100 ng/mL, 24h). 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 1 hour at room temperature. The cells were labeled with EPS8 Polyclonal Antibody (Product # PA5-21295) at 1:100 dilution 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 cytosolic localization of EPS8. Panel e represents untreated cells with no EPS8 expression. Panel f shows 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: EPS8 Polyclonal Antibody detects EPS8 protein at cytosol and nucleus on human breast carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human breast carcinoma. EPS8 Polyclonal Antibody (Product # PA5-21295) dilution: 1:500. 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 EPS8 was performed in HepG2 whole cell extracts using 5 µg of EPS8 Polyclonal Antibody (Product # PA5-21295). Samples were transferred to a membrane and probed with EPS8 Polyclonal Antibody as a primary antibody and an HRP-conjugated anti-Rabbit IgG was used as a secondary antibody.

PA5-21295