- PA5-28150 - Invitrogen Antibodies PDCD4 antibody

Sabry R, Williams M, Werry N, LaMarre J, Favetta LA
International journal of molecular sciences 2022 Jul 27;23(15)

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of PDCD4 using 30 µg of A) A431 and B) HeLa lysate. Samples were loaded onto a 10% SDS-PAGE gel and probed with a PDCD4 polyclonal antibody (Product # PA5-28150) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of PDCD4 in Mouse tissue extracts (50 µg). Samples was separated by 10% SDS-PAGE and the membrane was probed with PDCD4 Polyclonal antibody (Product # PA5-28150) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using PDCD4 Polyclonal Antibody (Product # PA5-28150). Various whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with PDCD4 Polyclonal Antibody (Product # PA5-28150) diluted at 1:1,000. 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: PDCD4 Polyclonal Antibody detects PDCD4 protein by western blot analysis. Mouse tissue extracts (50 µg) was separated by 10% SDS-PAGE, and the membrane was blotted with PDCD4 Polyclonal Antibody (Product # PA5-28150) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-PDCD4 Polyclonal Antibody (Product # PA5-28150) and a 51 kDa band corresponding to Programmed cell death protein 4 was observed across the cell lines and tissues tested. Whole cell extracts (30 µg lysate) of SH-SY5Y(Lane 1), HeLa (Lane 2), HeLa treated with 1 1 µg/ml of Staurosporine for 2 hrs (Lane 3) and Mouse Thymus (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) 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 Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of PDCD4 in paraformaldehyde-fixed HeLa cells using a PDCD4 polyclonal antibody (Product # PA5-28150) (Green) at a 1:500 dilution. Alpha-tubulin filaments were labeled with Product # PA5-29281 (Red) at a 1:2000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Programmed cell death protein 4 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with PDCD4 Polyclonal Antibody (Product # PA5-28150) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), 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 dilution). Panel d represents the merged image showing nuclear and 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: Relative protein expression of PDCD4 & PTEN after miR-21 inhibition . Western blots ( A ) and graphical representations of PDCD4 ( B ) and PTEN ( C ) revealed that miR-21 inhibition significantly increased PDCD4 protein levels but did not affect PTEN. Transfections were done with LNA inhibitor probes at 0.5 muM for 12 h. Densitometric analysis was performed relative to the loading control, beta-actin. PTEN representation contains lane rearrangement; full original PTEN blot is included in the Supplementary File (Figure S4) . Different letters indicate significant differences, with b indicating a significantly different mean than a at p < 0.005. Bars represent the mean +- SEM.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Relative protein expression of PDCD4 & PTEN after miR-21 inhibition and BPA treatment . Western blots ( A ) and graphical representations of PDCD4 ( B ) and PTEN ( C ) revealed that BPA decreased PDCD4 protein, but miR-21 LNA treatment increased it. Neither BPA nor miR-21 LNA treatment affected PTEN protein. Transfections were done with LNA inhibitor probes at 0.5 muM for 12 h followed by BPA treatment for another 12 h. Densitometric analysis was performed relative to the loading control, B-actin. Different letters indicate significant differences, with b indicating a significantly different mean than a at p < 0.05 and c indicating a significantly different mean than a and b at p < 0.05. ab indicates no differences between a or b . Bars represent the mean +- SEM.

PA5-28150