PA1-28875

antibody from Invitrogen Antibodies

Targeting: TFF1 BCEI, D21S21, HP1.A, HPS2, pNR-2, pS2

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-28875

Provider

Invitrogen Antibodies

Product name

TFF1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Recommended positive controls: About 70% of breast carcinomas show partial staining of tumor cells..

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

250 µL

Concentration

Conc. Not Determined

Storage

4° C, do not freeze

References

Submitted references

Gastric Proteins MUC5AC and TFF1 as Potential Diagnostic Markers of Colonic Sessile Serrated Adenomas/Polyps.

Khaidakov M, Lai KK, Roudachevski D, Sargsyan J, Goyne HE, Pai RK, Lamps LW, Hagedorn CH
American journal of clinical pathology 2016 Nov 1;146(5):530-537

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-TFF1 Polyclonal Antibody (Product # PA1-28875) and a 9kDa band corresponding to Trefoil factor 1 was observed in MCF7 but not in MDA-MB-231 or MCF10A cell lines as reported in the literature. 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), MCF 10A (Lane 7), MCF 10A treated with Protein Transport Inhibitor Cocktail (1X for 4h) (Lane 8) 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.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Trefoil factor 1 was performed using 70% confluent log phase MCF7 cells. 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 45 minutes at room temperature. The cells were labeled with TFF1 Polyclonal Antibody (Product # PA1-28875) at 1:100 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). Panel d represents the merged image showing cytoplasmic localization. Panel e represents MDA-MB-231 cells showing no expression of TFF1. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Image 1 Expression of MUC5AC and TFF1 in normal colon ( A ), hyperplastic polyps (HPs) ( B ), and sessile serrated adenomas/polyps (SSA/Ps) ( C ) (x10). Immunofluorescence and immunoperoxidase staining of the same region is shown for representative control (normal), HP, and SSA/P samples. As expected, the signals for gastric proteins MUC5AC and TFF1 were negligible in normal colonic mucosa. MUC5AC and TFF1 immunopositive cells were present in HPs. In SSA/Ps, MUC5AC and TFF1 were typically expressed by most of the epithelial cells along the entire length of architecturally compromised crypts. SSA/Ps, compared with HPs, also showed significant coexpression of both MUC5AC and TFF1 in merged immunofluorescence analyses.