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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Other assay [1]
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- Product number
- 712138 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IGF2BP1 Recombinant Polyclonal Antibody (18HCLC)
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with Monkey, Dog, Cat.
- Antibody clone number
- 18HCLC
- Concentration
- 0.5 mg/mL
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
Molecular therapy. Nucleic acids 2021 Jun 4;24:755-767
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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.
- 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.
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.
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.
