MA1-25323

antibody from Invitrogen Antibodies

Targeting: TMPO LAP2, LEMD4, TP

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-25323

Provider

Invitrogen Antibodies

Product name

LAP2 Monoclonal Antibody (6E10)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

Recommended positive controls: K562.

Antibody clone number

6E10

Concentration

2.3 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using LAP2 Monoclonal Antibody (6E10) (Product # MA1-25323) and a 38 kDa band corresponding to LAP2 was observed across cell lines and tissues tested along with an uncharacterized band (*) at ~60 kDa. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), NIH-3T3 (Lane 2), U-87 MG (Lane 3), HEK-293 (Lane 4), Mouse liver (Lane 5) and Rat liver(Lane 6) were electrophoresed using 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 (3ug/ml) and detected by chemiluminescence with 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 LAP2 was performed using 70% confluent log phase HeLa 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 LAP2 Monoclonal Antibody (6E10) (Product # MA1-25323) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (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, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin Immunoprecipitation (ChIP) assay of endogenous LAP2 protein using Anti-LAP2 Antibody: ChIP was performed using Anti-LAP2 Mouse monoclonal Antibody (Product # MA1-25323, 5 µg) on sheared chromatin from Hela cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Mouse IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to RAD51, DDB1 and MMP15 transcriptional start sites and SAT2 satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.