- PA5-31863 - Invitrogen Antibodies TFF1 antibody

Submitted references TFF-1 Functions to Suppress Multiple Phenotypes Associated with Lung Cancer Progression.

Minegishi K, Dobashi Y, Tsubochi H, Hagiwara K, Ishibashi Y, Nomura S, Nakamura R, Ohmoto Y, Endo S
OncoTargets and therapy 2021;14:4761-4777

Heparanase Promotes Syndecan-1 Expression to Mediate Fibrillar Collagen and Mammographic Density in Human Breast Tissue Cultured ex vivo.

Huang X, Reye G, Momot KI, Blick T, Lloyd T, Tilley WD, Hickey TE, Snell CE, Okolicsanyi RK, Haupt LM, Ferro V, Thompson EW, Hugo HJ
Frontiers in cell and developmental biology 2020;8:599

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Trefoil factor 1 using 30 µg of MCF-7 lysate. Samples were loaded onto a 15% SDS-PAGE gel and probed with a Trefoil factor 1 polyclonal antibody (Product # PA5-31863) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of TFF1 was performed by separating 30 µg of non-transfected (–) and transfected (+) 293T whole cell extracts by 15% SDS-PAGE. Proteins were transferred to a membrane and probed with a TFF1 Polyclonal Antibody (Product # PA5-31863) 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)
Main image
Experimental details: Western Blot analysis of TFF1 was performed by separating 30 µg of whole cell extracts by 15% SDS-PAGE. Proteins were transferred to a membrane and probed with a TFF1 Polyclonal Antibody (Product # PA5-31863) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-TFF1 Polyclonal Antibody (Product # PA5-31863) and a 9kDa band corresponding to Trefoil factor 1 was observed in MCF7 but not in the MDA-MB-231 cell line, as reported in the literature. Uncharacterized bands (*) were observed between 40-50 kDa. Whole-cell extracts (30 µg lysate) of MCF7 (Lane 1), MCF7 treated with Beta Estradiol (10 nanomolar for 24h) (Lane 2), MCF7 (Lane 3), MCF7 treated with Protein Transport Inhibitor Cocktail (1X for 4h) (Lane 4) MDA-MB-231 (Lane 5), MDA-MB-231 treated with Protein Transport Inhibitor Cocktail (1X for 4h) (Lane 6) were electrophoresed using Novex™ 16% Tricine Protein Gel (Product # EC6695BOX). 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 (1:500 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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). The expression of TFF1 was also found to be upregulated upon Beta Estradiol treatment.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Trefoil factor 1 in methanol-fixed MCF-7 cells using a Trefoil factor 1 polyclonal antibody (Product # PA5-31863) at a 1:500 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry (Paraffin) analysis of TFF1 was performed in paraffin-embedded mouse stomach tissue using TFF1 Polyclonal Antibody (Product # PA5-31863) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of paraffin-embedded human gastric cancer, using Trefoil factor 1 (Product # PA5-31863) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Expression of endogenous TFF-1 protein in cultured cells from human lung carcinoma. ( A ) Expression of the TFF-1 protein was detected at around 13kDa by immunoblotting analysis only in PC-3 cells. ( B ) Schematic presentation of TFF-1 expression and the amount secreted into the medium analyzed by ELISA. ( C ) Result of qRT-PCR analysis of TFF-1 mRNA expression, expressed relative to values obtained for H1975. ( B and C ) Square and bar indicate mean value and standard deviation, respectively, obtained from three replicated experiments with three replicate samples. Abbreviations : AC, adenocarcinoma; SCC, squamous cell carcinoma; LCC, large cell carcinoma; SmCC, small cell carcinoma; Marker, molecular weight marker.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 Establishment of TFF-1-overexpressing cell lines. SBC-5 and LC-AI cells were transfected with a vector containing the cDNA encoding TFF-1/FLAG (pBA-TFF1) or empty vector (pBA-neo). ( A ) Expression of this fusion protein was evaluated by immunoblotting analysis with anti-TFF-1 antibody. Expression was not observed in parental SBC-5 or LC-AI cells (BC), but a single band of around 15kDa was detected after transfection of pBA-TFF1 (G-TFF1) in 4 clones, but not observed in cells transfected with pBA-neo (G-neo). Expression was normalized to the beta-actin loading control. ""Marker"" indicates molecular weight marker. ( B and C ) Images of immunofluorescent staining in parental cells (BC), pBA-neo (G-neo) or pBA-TFF1-transfected cells (G-TFF1) of SBC-5 ( B ), LC-AI ( C ) cells are shown. Cells were stained with anti-TFF-1 antibody and with 4',6-diamidino-2-phenylindole (DAPI). Scale bar indicates 100mum. ( D ) Schematic presentation of TFF-1 mRNA expression after transfection with pBA-neo or pBA-TFF1 (3 clones; CL) expressed relative to values obtained for H1975. Square and bar indicate mean values and standard deviations, respectively, obtained by qRT-PCR analysis from three replicated experiments with three replicate samples. Results were statistically analyzed and p-values were obtained by Fisher's PLSD test.

PA5-31863

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