MA5-32102

antibody from Invitrogen Antibodies

Targeting: PLEC EBS1, PCN, PLEC1, PLTN

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Antibody Data

Product number

MA5-32102

Provider

Invitrogen Antibodies

Product name

Plectin Recombinant Rabbit Monoclonal Antibody (SY29-04)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Antibody clone number

SY29-04

Vial size

100 µL

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Plectin was performed by loading 10 µg of WT (lane 1) and PLEC CRISPR KO (lane 2) U2OS cell lysates in RIPA buffer onto a 4-15% gradient polyacrylamide gel. Proteins were transferred to nitrocellulose membrane and blocked in 5% milk. Ponceau stained transfer of blot is shown. Plectin was detected above 245 kDa using a Plectin recombinant monoclonal antibody (Product # MA5-32102) at a dilution of 1:5,000 in 5% BSA in TBST overnight at 4 deg, followed by secondary antibody diluted to 0.2 µg/mL using Goat anti-Rabbit IgG (H+L) HRP antibody (Product # 65-6120). Chemiluminescent detection was performed using Pierce ECL Western Blotting Substrate (Product # 32106). Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence of Plectin was performed using parental and Plectin CRISPR KO cells that were transfected with a GFP or mCherry plasmid, respectively. At 48 hrs post transfection parental and KO cells were mixed and plated to a 1:1 ratio on coverslips as a mosaic and incubated for 24 hrs. Cells were fixed in 4% PFA for 15 min and permeabilized with 0.1% Triton X-100. Cells were stained with Plectin recombinant monoclonal antibody (Product # MA5-32102) at a 1:1,000 dilution overnight at 4 deg. Secondary antibody incubation was performed using 1 µg/mL of Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Alexa Fluor 647 antibody (Product # A-21245) for 1 hr at RT. Imaging was performed with a 40X oil objective and analysis was performed using Image J. Cell image represents a single focal plane. Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.