- PA5-47030 - Invitrogen Antibodies TACSTD2 antibody

Submitted references SARS-CoV-2 can infect and propagate in human placenta explants.

Fahmi A, Brügger M, Démoulins T, Zumkehr B, Oliveira Esteves BI, Bracher L, Wotzkow C, Blank F, Thiel V, Baud D, Alves MP
Cell reports. Medicine 2021 Dec 21;2(12):100456

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis from lysates of NCI-N87 human gastric carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-human TROP-2 Antigen Affinity-purified Polyclonal Antibody (Product # PA5-47030) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody. A specific band was detected for TROP-2 at approximately 45-50 kDa (as indicated). This experiment was conducted under reducing conditions.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of TROP2 in NCI‚N87 human gastric carcinoma cell line. Samples were incubated in TROP2 polyclonal antibody (Product # PA5-47030) using a dilution of 1 µg/mL followed by a HRP-conjugated Anti-Goat IgG secondary antibody. A specific band was detected for TROP‚2 at approximately 45-50 kDa (as indicated). This experiment was conducted under reducing conditions.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti- TROP2 Polyclonal Antibody (Product # PA5-47030) and 35 kDa band corresponding to TACSTD2 was observed across all the cell lines tested except K-562 and SH-SY5Y which are reported to be negative and NIH/3T3 and Rat kidney which are of non-human origin. Whole cell extracts (30 µg lysate) of MCF-7 (Lane 1) and MCF-7 treated with 5-aza-2-deoxycytidine (2.5 µM for 48 hr) (Lane 2), MCF 10A (Lane 3), MDA-MB-231 (Lane 4), NIH/3T3 (Lane 5), K-562 (Lane 6), SH-SY5Y (Lane 7) and Rat kidney (Lane 8) were electrophoresed using Novex® 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 (1µg/ml) and detected by chemiluminescence with Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 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)
Main image
Experimental details: Immunofluorescence analysis of TROP2 was performed using 70% confluent log phase MCF-7 cells treated with 5-aza-2-deoxycytidine (2.5 µM for 48 hours). 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 TROP2 Polyclonal Antibody (Product # PA5-47030) at 2 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Rabbit anti-Goat IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A27012) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing increased TROP2 expression and localization to plasma membrane upon treatment with 5-aza-2-deoxycytidine. Panel e shows untreated cells with lower expression of TROP2. Panel f 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: Immunohistochemical analysis of TROP2 in immersion fixed paraffin-embedded sections of human brain (frontal cortex). Samples were incubated in TROP2 polyclonal antibody (Product # PA5-47030) using a dilution of 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic . Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown) and counterstained with hematoxylin (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometric analysis of PC-3 human prostate cancer cell line was stained with Goat Anti-human TROP-2 Antigen Affinity-purified Polyclonal Antibody (Product # PA5-47030, filled histogram) or control antibodyopen histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry of TROP2 in PC‚3 human prostate cancer cell line. Samples were incubated in TROP2 polyclonal antibody (Product # PA5-47030) or control antibody followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody.

PA5-47030

Antibody data