25-5370-41

antibody from Invitrogen Antibodies

Targeting: AREG AREGB, SDGF

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

25-5370-41

Provider

Invitrogen Antibodies

Product name

Anti-Amphiregulin Monoclonal Antibody (AREG559), PE-Cyanine7, eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The AREG858 monoclonal antibody reacts with human Amphiregulin (AR). AR is a member of the Epidermal Growth Factor (EGF) family and is expressed as a transmembrane precursor protein and released by proteolytic cleavage. Soluble AR is a ligand for the EGF receptor (EGFR) and the interaction promotes growth of normal epithelial cells, fibroblasts and keratinocytes. In the context of the immune system AR functions as an effector molecule for tissue repair and homeostasis, mediates resistance and tolerance to infection, and suppresses inflammation. Under different disease and activating conditions AR is expressed in T cells, group 2 innate lymphoid cells, basophils, mast cells, eosinophils, and neutrophils. Applications Reported: This AREG559 antibody has been reported for use in flow cytometric analysis. Applications Tested: This AREG559 antibody has been pre-diluted and tested by flow cytometric analysis of stimulated normal human peripheral blood cells using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resources Tab online. This may be used at 5 µL (0.5 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 488-561 nm; Emission: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

AREG559

Vial size

25 Tests

Concentration

5 µL/Test

Storage

4° C, store in dark, DO NOT FREEZE!

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Normal human peripheral blood lymphocytes were stimulated with Cell Stimulation Cocktail (plus protein transport inhibitors) (Product # 00-4975) (purple histogram) overnight or only with Protein Transport Inhibitor Cocktail (Product # 00-4980) (orange histogram). Cells were fixed and stained intracellularly with Anti-Human CD4 PerCP-Cyanine5.5 (Product # 45-0049-42) and Anti-Human Amphiregulin PE-Cy7 using the Intracellular Fixation and Permeabilization Buffers set (Product # 88-8824). Viable, CD4+ T cells in the lymphocyte gate, as determined by Fixable Viability Dye eFluor® 506 (Product # 65-0866), were used for analysis.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Normal human peripheral blood lymphocytes were stimulated with Cell Stimulation Cocktail plus protein transport inhibitors (Product # 00-4975) overnight. Cells were fixed and stained intracellularly with Anti-Human CD3 PerCP-Cyanine5.5 (Product # 45-0036-42) and Mouse IgG1 K Isotype Control PE-Cy7 (Product # 25-4714-82) (left) or Anti-Human Amphiregulin PE-Cy7 (Product # 25-5370) (right) using the Intracellular Fixation and Permeabilization Buffers set (Product # 88-8824). Viable cells in the lymphocyte gate, as determined by Fixable Viability Dye eFluor® 506 (Product # 65-0866), were used for analysis.