Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [2]
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Validation data
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- Product number
- PA5-17473 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IRAK-M Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 29 µg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of IRAK-M in extracts from HL-60 and Raji cell lines using IRAK-M polyclonal antibody (Product # PA5-17473).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-IRAK-M Polyclonal Antibody (Product # PA5-17473) and a 60 kDa band corresponding to Interleukin-1 receptor-associated kinase 3 was observed in THP-1 cells differentiated with PMA and in HEL 92.1.7 which are reported to be positive and not in other cell lines. Whole cell extracts (30 µg lysate) of THP-1 (Lane 1), THP-1 treated with PMA (200 nM for 6 hours) (Lane 2), HEL 92.1.7 (Lane 3), K-562 (Lane 4), Jurkat (Lane 5), Raji (Lane 6), BeWo (Lane 7) and SK-O-V3 (Lane 8) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) 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 Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of IRAK-M in THP-1 cells differentiated overnight with TPA, using an IRAK-M polyclonal antibody (Product # PA5-17473).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Interleukin-1 receptor-associated kinase 3 was performed using 70% confluent log phase THP-1 and THP-1 differentiated with PMA (200 nM for 6 hours). 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 IRAK-M Polyclonal Antibody (Product # PA5-17473) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), 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 Plasma membrane and Cytoplasmic localization. Panel e represents THP-1 cells having no expression of IRAK-M. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.