PA5-19302

antibody from Invitrogen Antibodies

Targeting: POU2F1 OCT1, OTF1

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-19302

Provider

Invitrogen Antibodies

Product name

OCT1 (POU2F1) Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with bovine, canine, mouse, porcine and rat based on sequence homology.

Reactivity

Human

Host

Goat

Isotype

IgG

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot staining of nuclear HeLa cell lysate using Product # PA5-19302 at a concentration of 0.5 µg/mL, the primary antibody incubation was 1 hour and the detection method was chemiluminescence.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of OCT1 was achieved by transfecting HeLa with OCT1 specific siRNAs (Silencer® select Product # s10848, s10847). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the OCT1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with OCT1 (POU2F1) Polyclonal Antibody (Product # PA5-19302, 0.5 µg/mL) and Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to OCT1.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using anti-OCT1 (POU2F1) Polyclonal Antibody (Product # PA5-19302) and ~80 kDa band corresponding to OCT1 was observed across the cell lines tested. An uncharacterized band (*) was observed at ~ 30 kDa. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), Ramos (Lane 2), HEK-293 (Lane 3) and HCT 116 (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 (0.5 µg/mL) and detected by chemiluminescence with Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).