Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
- Flow cytometry [1]
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- Product number
- MA5-32256 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- GAP43 Recombinant Rabbit Monoclonal Antibody (SC60-06)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- 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
- SC60-06
- 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 GAP43 in mouse brain lysate using a GAP43 Monoclonal antibody (Product # MA5-32256) at a dilution of 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockout of GAP43 was achieved by CRISPR-Cas9 genome editing. Western blot analysis of GAP43 was performed by loading 20 µg of U-87 MG wild type (Lane 1), U-87 MG CAS9 (Lane 2), U-87 MG GAP43 KO whole cell extracts. The blot was probed with Anti-GAP43 Rabbit Monoclonal Antibody (SC60-06) (Product # MA5-32256) using 1:1000 dilution and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036) using 1:4000 dilution. Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to GAP43.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-GAP43 Recombinant Rabbit Monoclonal Antibody (SC60-06)(Product # MA5-32256) and a 48kDa band corresponding to GAP43 was observed across cell lines and tissues tested except in A549, A-431 and Mouse Lung which are reported to be low to negative for GAP43 expression. An increase in GAP43 expression was observed in SH-SY5Y on neuronal differentiation. Membrane enriched extracts (30ug lysate) of SH-SY5Y (Lane 1), SH-SY5Y differentiated to neurons (Lane 2), SK-N-SH (Lane 3), IMR-32 (Lane 4), A549 (Lane 5), A-431 (Lane 6) and tissue lysates of Mouse Lung (Lane 7), Mouse Brain (Lane 8), Rat Brain (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 (1:2000 dilution) and detected by chemiluminescence with 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).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of GAP43 in Hela cells using a GAP43 Monoclonal antibody (Product # MA5-32256) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of GAP43 in SHG-44 cells using a GAP43 Monoclonal antibody (Product # MA5-32256) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of GAP43 of paraffin-embedded Mouse brain tissue using a GAP43 Monoclonal antibody (Product #MA5-32256). Counter stained with hematoxylin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometric analysis of GAP43 in SH-SY-5y cells using a GAP43 Monoclonal Antibody (Product # MA5-32256) at a dilution of 1:50, as seen in red compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.