MA5-15068

antibody from Invitrogen Antibodies

Targeting: IRS1 HIRS-1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-15068

Provider

Invitrogen Antibodies

Product name

IRS1 Monoclonal Antibody (K.346.8)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

It is not recommended to aliquot this antibody.

Antibody clone number

K.346.8

Concentration

84 µg/mL

References

Submitted references

Berberine induces lipolysis in porcine adipocytes by activating the AMP‑activated protein kinase pathway.

Yang Y, Lu R, Gao F, Zhang J, Liu F
Molecular medicine reports 2020 Jun;21(6):2603-2614

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-IRS1 Monoclonal Antibody (K.346.8) (Product # MA5-15068) and a ~130kDa band corresponding to Insulin receptor substrate 1 was observed across cell lines tested . Whole cell extracts (50 µg lysate) of MIA PaCa-2 (Lane 1), PANC-1 (Lane 2) were electrophoresed using 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 (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20000 using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).Relative expression observed between MIA Paca-2 and Panc-1.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Insulin receptor substrate 1 was performed using 70% confluent log phase MIA PaCa-2 cells. The cells were fixed with 4% paraformaldehyde for 15 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 IRS1 Monoclonal Antibody (K.346.8) (Product # MA5-15068) 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), 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 withRhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nucleus and cytoplasm localization. Panel e represents Panc-1. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60x magnification.