Antibody data
- Antibody Data
- Antigen structure
- References [2]
- Comments [0]
- Validations
- Western blot [2]
- ELISA [1]
- Other assay [2]
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Validation data
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- Product number
- MA1-088 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IGF1 Monoclonal Antibody (1C5-1A2)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- MA1-088 (1C5-1A2) has been successfully used as a coating antibody in a sandwich ELISA with Product # PA1-130 as the detection antibody.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 1C5-1A2
- Vial size
- 200 µg
- Concentration
- 1 mg/mL
- Storage
- -20°C
Submitted references Structural basis for assembly and disassembly of the IGF/IGFBP/ALS ternary complex.
Cross-Linking/Mass Spectrometry Uncovers Details of Insulin-Like Growth Factor Interaction With Insect Insulin Binding Protein Imp-L2.
Kim H, Fu Y, Hong HJ, Lee SG, Lee DS, Kim HM
Nature communications 2022 Jul 30;13(1):4434
Nature communications 2022 Jul 30;13(1):4434
Cross-Linking/Mass Spectrometry Uncovers Details of Insulin-Like Growth Factor Interaction With Insect Insulin Binding Protein Imp-L2.
Pompach P, Viola CM, Radosavljević J, Lin J, Jiráček J, Brzozowski AM, Selicharová I
Frontiers in endocrinology 2019;10:695
Frontiers in endocrinology 2019;10:695
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of human IGF1 was performed by loading the indicated amounts of recombinant human IGF1 protein per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with StartingBlock (TBS) Blocking Buffer (Product # 37542) for at least 1 hour. The membrane was probed with an IGF1 monoclonal antibody (Product # MA1-088) at a concentration of 1 µg/mL overnight at 4C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of human IGF1 was performed by loading the indicated amounts of recombinant human IGF1 protein per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with StartingBlock (TBS) Blocking Buffer (Product # 37542) for at least 1 hour. The membrane was probed with an IGF1 monoclonal antibody (Product # MA1-088) at a concentration of 1 µg/mL overnight at 4C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Sandwich ELISA analysis of an anti-human IGF1 monoclonal antibody (Product # MA1-088) was performed by loading 100 µL per well of antibody (Product # MA1-088) at a concentration of 1 µg/mL overnight at room temperature. The plate was washed 3 times with ELISA Wash Buffer (Product # N503), and 100 µL of recombinant human IGF1 was added to wells in duplicate at 10, 5, 2.5, 1.25, 0.625, 0.312 and 0 ng/mL concentrations and the samples were incubated for 2 hours at room temperature. The plate was washed, then incubated with 100 µL per well of an IGF1 polyclonal antibody (Product # PA1-130), biotinylated using EZ-Link Sulfo-NHS-LC-Biotinylation Kit (Product # 21435). The biotinylated antibody was loaded at a concentration of 2.0 µg/mL (red line), 1.0 µg/mL (blue line), and 0.5 µg/mL (green line) for 1 hour at room temperature, followed by 100 µL per well of Ultra Streptavidin-HRP (Product # N504) at a dilution of 1:10,000 for 30 minutes at room temperature. Detection was performed by adding 100 µL of 1-Step Ultra TMB substrate (Product # 34028) per well and incubating for 20 minutes at room temperature in the dark. The plate was then stopped with 100 µL per well of 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 4 SDS-PAGE and western blot analysis of photocrosslinked IGF-1 or des(63-70)-IGF-1 to Imp-L2. Imp-L2 (line 1) was photocrosslinked to SDA-modified IGF-1 (line 2) or des(63-70)-IGF-1 (line 3). Samples were resolved by SDS-PAGE on 14% gels and stained either by Coomassie blue or blotted to PVDF membrane and detected with anti-IGF-1 antibody (1C5-1A2) (MA1-088, ThermoFisher). Molecular weight standards (BioRad) are shown in line 4.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Release of CBP3 from the ternary complex after CLD3 proteolysis. a Fluorescence-detection size-exclusion chromatography (FSEC) of binary or ternary complexes after 10 h proteolysis by thrombin. The elution profile for standard IGF1-GFP (yellow), IGF1-GFP-strep/IGFBP3 binary complex (blue), and IGF1-GFP/IGFBP3/ALS-His ternary complex (red) are indicated as a control (top). b Immunoblot analysis of IGF1/myc-IGFBP3-Strep/ALS complex after 10 h proteolysis by thrombin and subsequent SEC. Anti-myc and anti-Strep antibody were used to detect NBP3 and CBP3, respectively. No-thrombin treatment served as negative control (Ctrl). c FSEC of the ternary complex (IGF1/IGFBP3-GFP/ALS-His) after thrombin digestion for indicated time. Fluorescence signal (GFP) was monitored to examine the dissociation of CBP3 from the ternary complex. d Quantification of immunoblot analysis for IGF1R phosphorylation after treatment of intermediate ternary complex. HEK293A cells were treated with IGF1-His (5 nM, positive control), IGF1-His/IGFBP3/ALS ternary complex (5 nM, none), or intermediate ternary complex (5 nM). The intermediate ternary complex was prepared by protease digestion (thrombin, ADAM12 and PAPP-A2) and subsequent SEC purification. No treatment was used for negative control (Ctrl). Data from three independent experiments ( n = 3) were analyzed and relative pIGF1R/IGF1R (Fold to Ctrl) were expressed as mean +- SD (ns, not significant vs. ctrl; **** P < 0.0001 vs. ctrl; #### P < 0.0001 vs. none