MA1-82995

antibody from Invitrogen Antibodies

Targeting: AGT SERPINA8

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-82995

Provider

Invitrogen Antibodies

Product name

Angiotensin I Monoclonal Antibody (BGN/KA/4H)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This antibody does not react with angiotensin II, III, or angiotensinogen in ELISA.

Antibody clone number

BGN/KA/4H

Concentration

1 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Angiotensinogen Mouse Monoclonal Antibody (Product # MA1-82995) and a 53 kDa band corresponding to Angiotensinogen was observed. Whole cell extracts (30 µg lysate) of MDA-MB-231 (Lane 1), MDA-MB-231 treated with Protein Transport Inhibitor (1X for 4 Hours) (Lane 2), MDA-MB-231 treated with 1,4,5,6,7,8-Hexahydro-2,7,7-trimethyl-4-(4-nitrophenyl)-5-oxo-3-quinolinecarboxylic acid cyclohexyl ester (Lane 3), HepG2 (Lane 4) HepG2 treated with Protein Transport Inhibitor (Lane 5) and HepG2 treated with 1,4,5,6,7,8-Hexahydro-2,7,7-trimethyl-4-(4-nitrophenyl)-5-oxo-3-quinolinecarboxylic acid cyclohexyl ester (Lane 6) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). 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 (2 µg/mL) and detected by chemiluminescence Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Angiotensinogen was performed using 70% confluent log phase HepG2 cells treated with 1X protein transport inhibitor (Product # 00-4980-03) for 4 hours. 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 1 hour at room temperature. cells were labeled with Angiotensinogen Monoclonal Antibody (Product # MA1-82995) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A28175) at a dilution of 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 with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing ER membrane/cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.