Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Immunohistochemistry [3]
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Validation data
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- Product number
- PA5-19969 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IRAK-M Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- In Western blot applications, this antibody detects a band at ~68kDa. A suggested positive control is mouse spleen tissue lysate. PA5-19969 can be used with blocking peptide PEP-0094.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 1 mg/mL
- Storage
- Maintain refrigerated at 2-8°C for up to 3 months. For long term storage store at -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of mouse spleen (M) and rat liver (R) tissue lysates using a IRAK-M polyclonal antibody (Product # PA5-19969) at 1 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of IRAK-M in (M) mouse spleen and (R) rat liver tissue lysates with IRAK-M Polyclonal Antibody (Product # PA5-19969) at 1 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-IRAK-M Polyclonal Antibody (Product # PA5-19969) and a 60 kDa band corresponding to Interleukin-1 receptor-associated kinase 3 was observed in THP-1 cells differentiated with PMA and not in undifferentiated THP-1 cells. Whole cell extracts (30 µg lysate) of THP-1 (Lane 1) and THP-1 treated with PMA (200 nM for 6 hours) (Lane 2) 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 µg/mL) 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).An uncharacterized band of ~50 kDa was also observed in differentiated THP-1 cells.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence of IRAK2 in A20 cells with IRAK-M Polyclonal Antibody (Product # PA5-19969) at 20 µg/mL. Red: IRAK-M Blue: DAPI staining
- 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 # PA5-19969) at 10 µg/mL 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.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence of IRAK-M in Rat Liver tissue with IRAK-M Polyclonal Antibody (Product # PA5-19969) at 10 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical staining of rat liver tissue using IRAK-M Polyclonal Antibody (Product # PA5-19969) at 2 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry of IRAK-M in human spleen tissue with IRAK-M Polyclonal Antibody (Product # PA5-19969) at 5 µg/mL.