712138

antibody from Invitrogen Antibodies

Targeting: IGF2BP1 IMP-1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Gel shift (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

712138

Provider

Invitrogen Antibodies

Product name

IGF2BP1 Recombinant Polyclonal Antibody (18HCLC)

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Antibody clone number

18HCLC

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

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

References

Submitted references

Testicular orphan receptor 4 (TR4) promotes papillary thyroid cancer invasion via activating circ-FNLA/miR-149-5p/MMP9 signaling.

Ouyang X, Feng L, Yao L, Xiao Y, Hu X, Zhang G, Liu G, Wang Z
Molecular therapy. Nucleic acids 2021 Jun 4;24:755-767

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of IGF2BP1 was achieved by transfecting Hep G2 cells with IGF2BP1 specific siRNA (Silencer® select Product # s20916 & s20918). Western blot analysis (Fig a) was performed using whole cell extracts from IGF2BP1 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Anti-IGF2BP1 Recombinant Rabbit Polyclonal Antibody (Product # 712138, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this Western blot is shown in the histogram (Fig b). Loss of signal upon siRNA mediated knockdown 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 Hep G2 (Lane 1), PANC-1 (Lane 2), HeLa (Lane 3), IMR-32 (Lane 4), HT-29 (Lane 5), U-2 OS (Lane 6), NTERA-2 (Lane 7), NCCIT (Lane 8), F9 (Lane 9) and tissue extracts of Mouse Testis (Lane 10) and Mouse Liver (Lane 11). The blot was probed with Anti-IGF2BP1 Recombinant Rabbit Polyclonal Antibody (Product # 712138, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). An ~63 kDa band corresponding to IGF2BP1 was observed in relevant cell lines and tissues tested.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis,HepG2 cells were fixed and permiabilized detection of endogenous IGF2BP1 using Anti-IGF2BP1 Recombinant Rabbit Polyclonal Antibody (Product # 712138, 1:100) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Nuclei (blue) were stained using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Panel a-d) clearly demonstares cytoplasmic localization of IGF2BP1 in HepG2 cells . Panel e-h) shows no specific staining in HT29 cells which is IGF2BP1 negative,demonstrating specificity. The images were captured at 60X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Detection of binding of endogenous IGF2BP1 protein to specific RNA using Anti-IGF2BP1 Antibody: RNA Immunoprecipitation (RIP) was performed using Anti-IGF2BP1 Recombinant Rabbit Polyclonal Antibody (Product # 712138, 4 µg) on clarified whole cell lysate from 2 million Hep G2 cells. Normal Rabbit IgG was used as a negative IP control. Immunoprecipitated RNA was purified by RiboPure™ RNA Purification Kit (Product # AM1924) and analyzed by RT-PCR using the Power SYBR® Green RNA-to-CT™ 1-Step Kit (Product # 4389986) with primer pairs over IGF2, MYC, ACTB mRNA (positive) and 18s rRNA (negative). Data is presented as the fraction of immunoprecipitated RNA (%IP) normalized to the total amount of RNA used for immunoprecipitation.