PA5-19969

antibody from Invitrogen Antibodies

Targeting: IRAK3 IRAK-M

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-19969

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.

Concentration

1 mg/mL

Western blot

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.

Supportive validation

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.

Supportive validation

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.

Immunocytochemistry

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

Supportive validation

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.

Immunohistochemistry

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.

Supportive validation

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.

Supportive validation

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.