PA5-23968

antibody from Invitrogen Antibodies

Targeting: IGF2BP1 IMP-1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-23968

Provider

Invitrogen Antibodies

Product name

IGF2BP1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with chicken, mouse, rat and Xenopus and zebrafish based on sequence homology.

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Vial size

400 µL

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis using an IGF2BP1 polyclonal antibody (Product # PA5-23968) in K562 cell lysates (35 µg per lane).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of IGF2BP1 was achieved by transfecting Hep G2 with IGF2BP1 specific siRNAs (Silencer® select Product # s20916; s20918). Western blot analysis (Fig. a) was performed using whole cell extracts from the IGF2BP1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with IGF2BP1 Polyclonal Antibody (Product # PA5-23968, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to IGF2BP1.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of K-562 (Lane 1), IMR32 (Lane 2), Caco-2 (Lane 3), Hep G2 (Lane 4) and U-2 OS (Lane 5). The blot was probed with Anti-IGF2BP1 Polyclonal Antibody (Product # PA5-23968, 1:1000 dilution) and detected by chemiluminescence Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 63kDa band corresponding to IGF2BP1 was observed across all the cell lines tested.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of IGF2BP1 was performed using 70% confluent log phase Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with anti-IGF2BP1 Polyclonal Antibody (Product # PA5-23968) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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 and Cytoplasmic localization. Panel e 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 analysis in formalin-fixed, paraffin-embedded human prostate carcinoma using an IGF2BP1 polyclonal antibody (Product # PA5-23968), followed by HRP-conjugated secondary antibody and DAB staining.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of K562 cells using an IGF2BP1 polyclonal antibody (Product # PA5-23968) (right) compared to a negative control cell (left) at a dilution of 1:10-50, followed by a FITC-conjugated goat anti-rabbit antibody