720266

antibody from Invitrogen Antibodies

Targeting: PCK1 PEPCK-C

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

720266

Provider

Invitrogen Antibodies

Product name

PCK1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Other

Description

These Polyclonal antibodies are of rabbit origin developed by immunizing animals with proteins or peptides. The polyclonal antibody is purified by affinity purification from the rabbit sera generated after immunizing the rabbits with a specific type of protein or peptide. The purified antibody is tested for its functionality in various relevant research applications. The antibody is developed for Research Use Only and is non-hazardous or non-infectious in nature.

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-PCK1 Polyclonal Antibody (Product # 720266) and a 34 kDa band corresponding to PCK1 was observed across the panel tested. Whole cell extracts (30 µg lysate) of MCF-7 (Lane 1), MDA-MB-231 (Lane 2), Hep G2 (Lane 3), HEK-293 (Lane 4), Caco-2 (Lane 5), HCT 116 (Lane 6) and Mouse Liver (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on Tissue extracts (30 µg lysate) of Mouse Liver (Lane 1), Mouse Kidney (Lane 2), Rat Kidney (Lane 3) and Membrane enriched cell extracts of LnCap (Lane 4), MDA-MB 231 (Lane 5) and MCF-7 (Lane 6). The blots were probed with Anti-PCK1/phosphoenolpyruvate carboxykinase Rabbit Polyclonal Antibody (Product # 720266, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 69 kDa and 34 kDa bands corresponding to two isoforms of PCK1 was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), 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® 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).