PA5-79600

antibody from Invitrogen Antibodies

Targeting: LIF CDF, DIA, HILDA

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-79600

Provider

Invitrogen Antibodies

Product name

LIF Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

500 µg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of LIF in recombinant human LIF protein using 0.5 ng per well. Sample was incubated with LIF polyclonal antibody (Product # PA5-79600) at a dilution of 0.5 µg/mL.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of LIF in, Lane 1: human Hela whole cell lysates, Lane 2: human placenta tissue lysates, Lane 3: human A431 whole cell lysates, Lane 4: human A549 whole cell lysates. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 µg of sample under reducing conditions. After Electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. The membrane was blocked with 5% non-fat milk/TBS for 1. 5 hour at RT. The membrane was incubated with LIF Polyclonal Antibody (Product # PA5-79600) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0. 1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5,000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit. A specific band was detected for LIF at approximately 36 kDa. The expected band size for LIF is at 22 kDa.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of LIF was performed using 80% confluent log phase MDA-MB-231 and T-47D, control and PTI treated, cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with LIF Polyclonal Antibody (Product # PA5-79600) at 1:100 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), (1:2500 dilution), for 45 minutes at room temperature (Panel a,e,i: Green). Nuclei (Panel b,f,j: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c,g,k: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image of untreated MDA-MB-231 cells with faint cytoplasmic staining for LIF that is enhanced upon PTI treatment (Panel h). Panel l represents T-47D treated with PTI showing no staining for LIF. The images were captured at 60X magnification. This difference in expression of LIF between MDA-MB-231 and T-47D is reported in several studies and is shown to influence the tumorigenesis and metastatic potential of these cancer cells [doi: 10.18632/oncotarget.1772; 10.18632/oncotarget.6756; 10.1158/1535-7163.MCT-18-1258]

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of LIF on paraffin-embedded human lung cancer tissue. Sample was incubated with LIF polyclonal antibody (Product# PA5-79600).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of LIF on paraffin-embedded human placenta tissue. Sample was incubated with LIF polyclonal antibody (Product# PA5-79600).