PA5-76633
antibody from Invitrogen Antibodies
Targeting: OPTN
FIP-2, FIP2, GLC1E, HIP7, HYPL, NRP, TFIIIA-INTP
Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [1]
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Validation data
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- Product number
- PA5-76633 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Optineurin Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen and the purity is > 95% (by SDS-PAGE).
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- 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.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Optineurin. Samples were incubated with Optineurin polyclonal antibody (Product # PA5-76633).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Optineurin was performed by loading 30 µg of HCT 116 (Lane 1) and HCT 116 - Optineurin knockout (Lane 2) whole cell lysate. The blot was probed with Anti-Optineurin Polyclonal Antibody (Product # PA5-76633, 1:1,000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4,000 dilution). Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to Optineurin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Optineurin Polyclonal Antibody(Product # PA5-76633) and a 60kDa band corresponding to Optineurin was observed across in all tested cell lines and tissues, except Mouse Eye, Mouse Liver, Mouse Kidney and Rat Eye. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), A-431 (Lane 2), HEK-293 (Lane 3), Mouse Retina (Lane 4), Mouse Eye (Lane 5), Mouse Liver (Lane 6), Mouse Kidney (Lane 7), Rat Retina (Lane 8), Rat Eye (Lane 9) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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 (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Optineurin was performed using 70 percent confluent log phase A-431 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 45 minutes at room temperature. The cells were labeled with Optineurin Polyclonal Antibody (Product # PA5-76633) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (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 cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.