MA5-16418
antibody from Invitrogen Antibodies
Targeting: IKZF1
hIk-1, Hs.54452, IKAROS, LyF-1, PPP1R92, ZNFN1A1
Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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Validation data
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- Product number
- MA5-16418 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-IKAROS Monoclonal Antibody (SP108)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- Heat-mediated antigen retrieval is recommended prior to staining, using an EDTA buffer, pH 8.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 10 min at room temperature. A suggested positive control is HeLa cell lysate.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- SP108
- Vial size
- 500 µL
- Storage
- 4° C, do not freeze
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-IKAROS Monoclonal Antibody (SP108) (Product # MA5-16418) and 60, 52 and 40 kDa bands corresponding to IKAROS isoforms were observed across relevant cell lines, tissues tested except in low expressing A-431 and Mouse Mammary Gland tissue. Nuclear enriched extracts (40 µg lysate) of Reh (Lane 1), Jurkat (Lane 2), Ramos (Lane 3), MOLT-4 (Lane 4), THP-1 (Lane 5), A-431 (Lane 6), Mouse Thymus (Lane 7), Mouse Mammary gland (Lane 8) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 IKAROS was performed using 70% confluent log phase Jurkat and A-431 cells. The cells were fixed with 4% paraformaldehyde for 5 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with IKAROS Monoclonal Antibody (SP108) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then 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). 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 nuclear localization. Panel e represents A-431 cells having no expression of IKAROS. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of IKAROS using anti-IKAROS Monoclonal Antibody (Product # MA5-16418) in Tonsil Tissue. The recommened dilution for this antibody in immunohistochemistry applications is 1:100.