PA5-16621

antibody from Invitrogen Antibodies

Targeting: AREG AREGB, SDGF

Western blot (Supportive data in Antibodypedia)

Immunoprecipitation (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-16621

Provider

Invitrogen Antibodies

Product name

Amphiregulin Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

PA5-16621 targets Amphiregulin in IP and WB applications and shows reactivity with Human and mouse samples. The PA5-16621 immunogen is a synthetic 19-mer peptide, corresponding to aa 26-44 from the secreted amphiregulin of human origin.

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Vial size

500 µL

Concentration

1 mg/mL

Storage

4°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of MDA-MB-231 (Lane 1), MDA-MB-231 treated with PMA (100 ng PMA for 16h) (Lane 2), Caco-2 (Lane 3), A-431 (Lane 4), PC-3 (Lane 5), U-2 OS (Lane 6) and HCT-116 (Lane7).The blots were probed with Amphiregulin Rabbit polyclonal Antibody (Product # PA5-16621, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 25 kDa band corresponding to Amphiregulin was observed across the cell lines tested and was enhanced upon treatment in MDA-MB-231 cell line. Apart from desired band of 25 KDa, ~60 kDa band is seen across the cell lines showing the glycosylated form of the protein. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot of Amphiregulin using Amphiregulin Polyclonal Antibody (Product # PA5-16621) on LS174T Cells.

Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation of Amphiregulin using Amphiregulin Polyclonal Antibody (Product # PA5-16621) on Native Human LS174T Cells.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of Amphiregulin was done on MDA-MB-231 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Amphiregulin Rabbit Polyclonal Antibody (PA5-16621, red histogram) or with rabbit isotype control (pink histogram) at 3-5 µg/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control..

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation of Amphiregulin using Amphiregulin Polyclonal Antibody (Product # PA5-16621) on Native Human LS174T Cells.