PA5-79579

antibody from Invitrogen Antibodies

Targeting: LAMC2 BM600-100kDa, EBR2, EBR2A, kalinin-105kDa, LAMB2T, LAMNB2, nicein-100kDa

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-79579

Provider

Invitrogen Antibodies

Product name

Laminin gamma-1,2,3 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody reacts will all 3 isoforms of Laminin gamma: Laminin gamma-1, gamma-2 and gamma-3.

Concentration

500 µg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Laminin in Lane 1: HeLa whole cell lysate, Lane 2: HEPG2 whole cell lysate using 40-50 µg per well. Sample was incubated with Laminin (Product # PA5-79579) at a dilution of 0.5 µg/mL.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using anti-Laminin gamma-1,2,3 Polyclonal Antibody (Product # PA5-79579) and a 210 kDa band corresponding to Laminin gamma-1,2,3 was observed across the cell lines and tissues tested. Whole cell extracts (1% SDS) (30 µg lysate) of Hep G2 (Lane 1), HUVEC (Lane 2), A-431 (Lane 3), K-562 (Lane 4), PC-3 (Lane 5), Mouse Liver (Lane 6), Rat Liver (Lane 7), Mouse Lung (Lane 8), Rat Lung (Lane 9) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gels (Product # EA0375BOX). 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 Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25 µg/mL, 1:10000 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 Laminin gamma-1,2,3 was performed using 70% confluent log phase Hep G2 cells. 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. The cells were labeled with Laminin gamma-1,2,3 Polyclonal Antibody (Product # PA5-79580) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents K-562 cells showing no expression of Laminin gamma-1,2,3. Panel f shows control Hep G2 cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of Laminin on paraffin-embedded human lung cancer tissue. Sample was incubated with Laminin polyclonal antibody (Product# PA5-79579).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of Laminin in frozen section of human placenta tissues. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti-Laminin antibody (Product # PA5-79579) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.