MA5-27480

antibody from Invitrogen Antibodies

Targeting: IGF2BP3 CT98, IMP-3, IMP3

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-27480

Provider

Invitrogen Antibodies

Product name

IGF2BP3 Monoclonal Antibody (OTI4H5)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Reactivity

Human, Mouse, Rat

Host

Mouse

Isotype

IgG

Antibody clone number

OTI4H5

Vial size

100 µL

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of IGF2BP3 in HEK293T cells in untransfected (Left lane) and transfected (Right lane) samples using 5 µg per lane. The samples were separated by SDS-PAGE and probed with IGF2BP3 (Product # MA5-27480) monoclonal antibody.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-IGF2BP3 Mouse Monoclonal Antibody (Product # MA5-27480) and a 64 kDa band corresponding to IGF2BP3 was observed in cell lines and tissues tested except MCF7, SK-BR-3, Mouse Lung and Mouse Spleen as reported. Whole cell extracts (30 µg lysate) of SH-SY5Y (Lane 1), IMR-32 (Lane 2), MDA-MB-231 (Lane 3), MCF7 (Lane 4), SK-BR-3 (Lane 5), Mouse Placenta (Lane 6), Mouse Testis (Lane 7), Rat Testis (Lane 8), Mouse Spleen (Lane 9) and Mouse Lung (Lane 10) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein 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-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). A 25 kDa band (*) corresponding to circulating tissue IgG, was observed in mouse tissues.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of IGF2BP3 was performed using IMR-32 and MCF7 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 IGF2BP3 Mouse Monoclonal Antibody (Product # MA5-27480) at 1:100 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green) in IMR-32 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 IMR-32 cells, which is a positive model for IGF2BP3 expression showing a cytoplasmic and nuclear localization. Panel e represents the merged image ofMCF7 cells, that are null for IGF2BP3 protein expression. 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

Immunohistochemistry was performed on paraffin-embedded human placenta tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a IGF2BP3 monoclonal antibody (Product # MA5-27480) at a dilution of 1:250.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry was performed on paraffin-embedded human testicle tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a IGF2BP3 monoclonal antibody (Product # MA5-27480) at a dilution of 1:250.