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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
- Immunocytochemistry [3]
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- Product number
- MA5-24855 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-KIF23 Recombinant Rabbit Monoclonal Antibody (2E4)
- Antibody type
- Other
- 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
- 2E4
- Vial size
- 100 µL
- 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 KIF23 in Lane 1: A549 whole cell lysate, Lane 2: HeLa whole cell lysate. Samples were incubated with KIF23 monoclonal antibody (Product # MA5-24855) at a dilution of 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using anti-KIF23 Monoclonal Antibody (Product # MA5-24855) and ~110, 100 kDa bands corresponding to two isoforms of KIF23 were observed. Whole cell extracts (30 µg lysate) of MCF 10A (Lane 1), MDA-MB-231 (Lane 2), MCF-7 (Lane 3), U-87 MG (Lane 4), BJ (Lane 5), A549 (Lane 6) and OVCAR-3 (Lane 7) were electrophoresed using Novex® NuPAGE® 4-12% % Bis-Tris 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 (1:1000 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
- Immunofluorescence analysis of KIF23 in HeLa cells (green). Samples were treated with paraformaldehyde, permeabilized with 0.25% Triton and PBS, and incubated with KIF23 monoclonal antibody (Product # MA5-24855).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of KIF23 in A549 cells (green). Samples were treated with paraformaldehyde, permeabilized with 0.25% Triton and PBS, and incubated with KIF23 monoclonal antibody (Product # MA5-24855).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of KIF23 was performed using 70% confluent log phase MCF 10A and MCF-7 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 KIF23 Monoclonal Antibody (Product # MA5-24855) at 1:200 dilution 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 (Product # A32790) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green) in A549 cells. 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 of MCF-7, which is a positive model for KIF23 expression showing a nuclear localization. Panel e represents the merged image of MCF 10A, that are null for KIF23 protein expression. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
