PA5-28639

antibody from Invitrogen Antibodies

Targeting: RTN4R NOGOR

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-28639

Provider

Invitrogen Antibodies

Product name

Nogo Receptor Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Recommended positive controls: HepG2.

Concentration

1 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using Nogo Receptor Polyclonal Antibody (Product # PA5-28639). Sample (30 µg of whole cell lysate). Lane A: Hep G2. 10% SDS PAGE. Nogo Receptor Polyclonal Antibody (Product # PA5-28639) diluted at 1:1,000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Nogo Receptor Polyclonal Antibody (Product # PA5-28639) and 80 kDa band corresponding to Nogo receptor was observed along with uncharacterized bands (*) across cell lines and tissues tested except Mouse Skeletal Muscle, Mouse and Rat Heart. Tissue extracts of Mouse Brain (Lane 1), Rat Brain (Lane 2), Mouse cerebellum (Lane 3), Rat cerebellum (Lane 4), Mouse skeletal Muscle (Lane 5), Mouse Heart (Lane 6), Rat Heart (Lane 7), whole cell extracts (30 µg lysate) of Hep G2 (Lane 8), IMR-32 (Lane 9) and SH-SY5Y (Lane 10) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0342BOX). Relative expression of Nogo receptor was observed in Mouse Brain, Rat Brain, Mouse cerebellum, and Rat cerebellum in higher levels in comparison to low levels in Mouse Skeletal Muscle, Mouse Heart, and Rat heart using Anti-Nogo receptor Polyclonal Antibody (Product # PA5-28639) in Western Blot. Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with primary antibody (1:1000 Dilution) and detected by chemiluminescence Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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 Nogo Receptor was performed using SH-SY5Y 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 PRKCI Rabbit Polyclonal Antibody (Product # PA5-28639) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 conjugate (Product # A32790) 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 composite image showing membrane staining of Nogo receptor in SH-SY5Y cells. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.