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Rabbit anti igf 1

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Rabbit anti-IGF-1 is a primary antibody that specifically binds to the insulin-like growth factor 1 (IGF-1) protein. IGF-1 is a hormone that plays a crucial role in cell growth and metabolism. This antibody can be used to detect and quantify IGF-1 levels in various biological samples.

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Lab products found in correlation

Rabbit anti e cadherin, by Abcam (2 mentions) Goat anti shh, by R&D Systems (2 mentions) Igf 1 standards, by Abcam (2 mentions) Goat antirabbit igg1 conjugated to hrp, by Southern Biotech (2 mentions) Tmb peroxidase substrate, by Rockland Immunochemicals (2 mentions) Automated stainer, by Leica (1 mentions) Mouse anti β actin, by Santa Cruz Biotechnology (1 mentions) Rabbit anti hgf, by Santa Cruz Biotechnology (1 mentions) Rabbit anti anxa2, by Santa Cruz Biotechnology (1 mentions) Vectastain abc kit, by Vector Laboratories (1 mentions) Rabbit anti ki67, by Abcam (1 mentions) Biotinylated goat anti rabbit secondary antibody, by Vector Laboratories (1 mentions) Rabbit anti c met, by Santa Cruz Biotechnology (1 mentions) Supersignal west dura extended duration substrate, by Thermo Fisher Scientific (1 mentions) Radioimmunoprecipitation assay buffer, by Cell Signaling Technology (1 mentions) Anti mouse β catenin, by Abcam (1 mentions) Fluorescence microscope, by Leica (1 mentions) Fluoromount, by Vector Laboratories (1 mentions) Vectastain elite abc peroxidase kit, by Vector Laboratories (1 mentions) Vector sg, by Vector Laboratories (1 mentions)

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IGF-1 (51 citations) Anti-IGF-1 (13 citations) Rabbit anti- (5 citations)

9 protocols using rabbit anti igf 1

1

Evaluation of AnxA2 and Its Phosphorylation

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Elution of AnxA2 (26 (link))and Western blot with rabbit anti-AnxA2, mouse anti-P-Y23-AnxA2, mouse anti-beta-actin, rabbit anti-HGF (all from Santa Cruz Biotechnology), rabbit anti-IGF-1(Abcam) or goat anti-Shh (R&D Systems) antibodies were described previously (12 (link)) Immunohistochemistry (IHC) staining for E-Cadherin and HGF was performed with rabbit anti-E-Cadherin (Abcam) and rabbit anti-HGF (Santa Cruz Biotechnology) antibodies using a standard protocol on an automated stainer from Leica Microsystems. IHC for SMA was performed as previously described (27 (link)). IHC for AnxA2 was conducted with mouse anti-AnxA2(Invitrogen), or mouse anti-P-Y23-AnxA2 antibodies, IHC for Shh with goat-anti-Shh (R&D), and IHC for IGF-1 with rabbit-anti-IGF-1(Abcam) antibodies as described previously (13 (link)). All IHC slides were analyzed and scored by a pathologist (A.L). The levels of secreted IGF-1 were measured by ELISA (R&D) following the manufacturer’s instructions.
Rucki A.A., Foley K., Zhang P., Xiao Q., Kleponis J., Wu A.A., Sharma R., Mo G., Liu A., Van Eyk J., Jaffee E.M, & Zheng L. (2016). Heterogeneous stromal signaling within the tumor microenvironment controls the metastasis of pancreatic cancer. Cancer research, 77(1), 41-52.
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2

Multimarker IHC Analysis of Signaling Pathways

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Immunohistochemistry (IHC) staining for Shh, c-Met, E-Cadherin, IGF-1 and Ki67 were performed by hand. Tissue blocks with poor quality were excluded from the study. The slides for all stainings were hydrated; antigen retrieval was performed in a pressure cooker with citrate buffer (pH 6.0) for Shh and Ki67, in a steamer with citrate buffer (pH 6.0) for c-Met and IGF-1 and with Tris-EDTA buffer (pH 9.0) for E-Cadherin. Then, the slides were blocked in peroxidase, avidin and biotin block sequentially. Goat-anti-Shh (R&D), rabbit-anti-c-Met (Santa Cruz), rabbit anti-IGF-1 (Abcam), rabbit anti-E-Cadherin (Abcam), or rabbit anti-Ki67 (Abcam) primary antibodies at 1:50 (Shh, c-Met, E-Cadherin), 1: 500 (Ki67) and 4µg/ml (IGF-1) were added, and the slides were incubated for 1 hour at room temperature. Then, rabbit anti-goat or goat anti-rabbit biotinylated secondary antibodies (Vector Laboratories) respectively, were added for 30 minutes at room temperature. The signal was amplified and detected using the ABC Vectastain kit (Vector Laboratories) according to the manufacturer’s instructions. The slides were developed using DAB and counterstained by hematoxylin. Lastly, the slides were dehydrated and mounted. All IHC slides were analyzed and scored by a pathologist. Immunofluorescence staining of c-Met and Shh was described in Supplementary Materials and Methods.
Rucki A.A., Xiao Q., Muth S., Chen J., Che X., Kleponis J., Sharma R., Anders R.A., Jaffee E.M, & Zheng L. (2017). Dual inhibition of Hedgehog and c-Met pathways for pancreatic cancer treatment. Molecular cancer therapeutics, 16(11), 2399-2409.
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3

IGF-1 Sandwich ELISA Protocol

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IGF-1 standards (Abcam, Cambridge, UK) and sera samples were diluted in coating buffer (15 mM Na2CO3, 35 mM NaHCO3, 3 mM NaH3, pH9.6) and allowed to bind to Costar (3590) high-binding EIA/RIA plates overnight. Subsequently, plates were washed 3 times in PBS-0.1% Tween-20 (PBST), blocked with 3% milk in PBST (PTM) for 1 h and incubated in primary antibody, rabbit anti-IGF-1 in PTM 1:2000 (Abcam, Cambridge, UK) for 2 h at 37 °C. Next, plates were washed 3 times in PBST and incubated in secondary antibody goat antirabbit IgG1 conjugated to HRP in PTM 1:4000 (Southern Biotechnology, Birmingham, AL) at 37 °C for 1 h. Plates were then washed in PBST, incubated in TMB peroxidase substrate (Rockland) for 10 min at room temperature, followed by stop solution (2 N H2SO4). Absorbance was read at 450 nm.
Park J., Yan G., Kwon K.C., Liu M., Gonnella P.A., Yang S, & Daniell H. (2019). Oral delivery of novel human IGF-1 bioencapsulated in lettuce cells promotes musculoskeletal cell proliferation, differentiation and diabetic fracture healing. Biomaterials, 233, 119591.
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4

Quantifying IGF1 Protein in Flushed Tibia

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Flushed tibia was used for protein extraction using 1 × radioimmunoprecipitation assay buffer (Cell Signaling). Same amount of protein was loaded on sodium dodecyl sulfate–polyacrylamide gel. After transfer, membrane was incubated for 1 h with 5% nonfat dry milk, then incubated at 4 °C for overnight with rabbit anti-IGF1 (Abcam) antibody, then incubated with anti-rabbit secondary antibody for 1 h at room temperature. Blots were developed with Super Signal West Dura Extended Duration Substrate (Thermo Scientific), then re-probed with actin antibody (Santa Cruz Biotechnology, Inc) for loading control.
Xiao L., Zhou Y., Bokoliya S., Lin Q, & Hurley M. (2022). Bone loss is ameliorated by fecal microbiota transplantation through SCFA/GPR41/ IGF1 pathway in sickle cell disease mice. Scientific Reports, 12, 20638.
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5

Immunohistochemistry and Immunofluorescence Protocols

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For immunohistochemistry analysis, the sections were deparaffinized and boiled in a citrate buffer solution. The sections were stained using standard methods with rabbit anti-CD31 (PECAM-1, Santa Cruz Biotechnology, Inc. 1 : 100) as the primary antibody. The sections were incubated using a Vectastain Elite ABC-Peroxidase kit (Vector Laboratories, Burlingame, CA, USA), visualized using a Vector SG (Vector Laboratories), and counterstained with nuclear fast solutions (Vector Laboratories). For immunofluorescence, the sections were deparaffinized and boiled in a citrate buffer solution. They were stained using standard methods with rabbit anti-IGF-1 (dilution 1 : 100, Abcam, Cambridge, UK) and mouse anti-β-catenin (dilution 1 : 100, Abcam) as the primary antibodies. The sections were incubated with fluorescein isothiocyanate-conjugated goat anti-mouse and Texas red-conjugated goat anti-rabbit secondary antibodies (dilution 1 : 100, Santa Cruz Biotechnology, Inc.). The sections were fixed with fluoromount (Vector Laboratories) and viewed under a fluorescence microscope (Leica Microsystems, Wetzlar, Germany).
Kim H.J., Choi E.W., Choi E.J., Kim H.S., Kim J., Cho G., Kim H., Na S., Shin J.H., Do S.H, & Park B.J. (2021). Non-thermal plasma promotes hair growth by improving the inter-follicular macroenvironment. RSC Advances, 11(45), 27880-27896.
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6

Immunohistochemical Analysis of IGF1 Expression

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The immunohistochemistry (IHC) assay was performed as previously described.6 Briefly, after deparaffinized, rehydrated, and antigen retrieval, the sections were incubated with rabbit anti‐IGF1 (Abcam) overnight at 4°C. The sections were incubated with a biotinylated secondary antibody and stained with hematoxylin. Following this, the slides were imaged using the ScanScope microscope (Olympus).
The IGF1 expression based on IHC assay was assessed as previously described7 and examined by two pathologists, who were blinded to the identity of specimens.
Yang L., Tan Z., Li Y., Zhang X., Wu Y., Xu B, & Wang M. (2020). Insulin‐like growth factor 1 promotes proliferation and invasion of papillary thyroid cancer through the STAT3 pathway. Journal of Clinical Laboratory Analysis, 34(12), e23531.
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7

Immunostaining of Nerve Tissue Sections

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The harvested L4-6 DRGs and sciatic nerve segment were fixed in 4% paraformaldehyde, embedded in 5% sucrose, and cut on a cryostat into 12-μm-thick sections, which were immunostained with primary antibodies: mouse anti-neurofilament (NF) 200 (1:400, Sigma), rabbit anti-IGF-1 (1:200, Abcam), and mouse anti-S100 β (1:100), followed by reaction with fluorescently-labeled secondary antibodies (1:400, Invitrogen). Images were taken under fluorescence microscopy (Leica, Germany).
Zhu H., Xue C., Yao M., Wang H., Zhang P., Qian T., Zhou S., Li S., Yu B., Wang Y, & Gu X. (2018). miR-129 controls axonal regeneration via regulating insulin-like growth factor-1 in peripheral nerve injury. Cell Death & Disease, 9(7), 720.
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8

IGF-1 Sandwich ELISA Protocol

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IGF-1 standards (Abcam, Cambridge, UK) and sera samples were diluted in coating buffer (15 mM Na2CO3, 35 mM NaHCO3, 3 mM NaH3, pH9.6) and allowed to bind to Costar (3590) high-binding EIA/RIA plates overnight. Subsequently, plates were washed 3 times in PBS-0.1% Tween-20 (PBST), blocked with 3% milk in PBST (PTM) for 1 h and incubated in primary antibody, rabbit anti-IGF-1 in PTM 1:2000 (Abcam, Cambridge, UK) for 2 h at 37 °C. Next, plates were washed 3 times in PBST and incubated in secondary antibody goat antirabbit IgG1 conjugated to HRP in PTM 1:4000 (Southern Biotechnology, Birmingham, AL) at 37 °C for 1 h. Plates were then washed in PBST, incubated in TMB peroxidase substrate (Rockland) for 10 min at room temperature, followed by stop solution (2 N H2SO4). Absorbance was read at 450 nm.
Park J., Yan G., Kwon K.C., Liu M., Gonnella P.A., Yang S, & Daniell H. (2019). Oral delivery of novel human IGF-1 bioencapsulated in lettuce cells promotes musculoskeletal cell proliferation, differentiation and diabetic fracture healing. Biomaterials, 233, 119591.
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9

Protein Extraction and Western Blot Analysis

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Total protein was extracted from the indicated cells with RIPA buffer containing protease inhibitors (Beyotime). The Western blot analysis was performed as previously described8 The first antibodies in this study included rabbit anti‐IGF1 (Abcam), mouse anti‐MMP2 (Abcam), mouse anti‐MMP9 (Abcam), rabbit anti‐STAT3 (CST), rabbit anti‐p‐STAT3 (CST), and mouse anti‐GAPDH (Abcam). The band density was analyzed using ImageJ (NIH).
Yang L., Tan Z., Li Y., Zhang X., Wu Y., Xu B, & Wang M. (2020). Insulin‐like growth factor 1 promotes proliferation and invasion of papillary thyroid cancer through the STAT3 pathway. Journal of Clinical Laboratory Analysis, 34(12), e23531.
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