Product: IGFBP3 Antibody
Catalog: AF6203
Description: Rabbit polyclonal antibody to IGFBP3
Application: WB IHC
Reactivity: Human, Mouse
Prediction: Pig, Bovine, Horse, Sheep, Dog
Mol.Wt.: 31kDa; 32kD(Calculated).
Uniprot: P17936
RRID: AB_2835084

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 100ul $280 In stock
 200ul $350 In stock

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Dog(100%)
Clonality:
Polyclonal
Specificity:
IGFBP3 Antibody detects endogenous levels of total IGFBP3.
RRID:
AB_2835084
Cite Format: Affinity Biosciences Cat# AF6203, RRID:AB_2835084.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

Acid stable subunit of the 140 K IGF complex; Binding protein 29; Binding protein 53; BP 53; BP53; Growth hormone dependent binding protein; IBP 3; IBP-3; IBP3; IBP3_HUMAN; IGF binding protein 3; IGF-binding protein 3; IGFBP 3; IGFBP-3; IGFBP3; Insulin Like Growth Factor Binding Protein 3; Insulin-like growth factor binding protein 3 precursor; Insulin-like growth factor-binding protein 3;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P17936 IBP3_HUMAN:

Expressed by most tissues. Present in plasma.

Description:
This gene is a member of the insulin-like growth factor binding protein (IGFBP) family and encodes a protein with an IGFBP domain and a thyroglobulin type-I domain. The protein forms a ternary complex with insulin-like growth factor acid-labile subunit (IGFALS) and either insulin-like growth factor (IGF) I or II.
Sequence:
MQRARPTLWAAALTLLVLLRGPPVARAGASSAGLGPVVRCEPCDARALAQCAPPPAVCAELVREPGCGCCLTCALSEGQPCGIYTERCGSGLRCQPSPDEARPLQALLDGRGLCVNASAVSRLRAYLLPAPPAPGNASESEEDRSAGSVESPSVSSTHRVSDPKFHPLHSKIIIIKKGHAKDSQRYKVDYESQSTDTQNFSSESKRETEYGPCRREMEDTLNHLKFLNVLSPRGVHIPNCDKKGFYKKKQCRPSKGRKRGFCWCVDKYGQPLPGYTTKGKEDVHCYSMQSK

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Xenopus
0
Zebrafish
0
Chicken
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P17936 As Substrate

Site PTM Type Enzyme
T7 Phosphorylation
N116 N-Glycosylation
N136 N-Glycosylation
S138 Phosphorylation P68400 (CSNK2A1)
S140 Phosphorylation P68400 (CSNK2A1)
S145 Phosphorylation
S148 O-Glycosylation
S148 Phosphorylation
S151 O-Glycosylation
S151 Phosphorylation
S153 O-Glycosylation
S155 O-Glycosylation
S156 O-Glycosylation
S156 Phosphorylation
T157 O-Glycosylation
S161 O-Glycosylation
S170 Phosphorylation
S183 Phosphorylation P78527 (PRKDC)
Y190 Phosphorylation
S192 O-Glycosylation
S194 Phosphorylation P68400 (CSNK2A1)
T195 O-Glycosylation
T197 Phosphorylation
N199 N-Glycosylation
S201 Phosphorylation
Y210 Phosphorylation
S231 Phosphorylation
Y246 Phosphorylation
T277 O-Glycosylation

Research Backgrounds

Function:

IGF-binding proteins prolong the half-life of the IGFs and have been shown to either inhibit or stimulate the growth promoting effects of the IGFs on cell culture. They alter the interaction of IGFs with their cell surface receptors. Also exhibits IGF-independent antiproliferative and apoptotic effects mediated by its receptor TMEM219/IGFBP-3R.

PTMs:

Phosphorylated by FAM20C in the extracellular medium.

Subcellular Location:

Secreted.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed by most tissues. Present in plasma.

Subunit Structure:

Interacts with XLKD1 (By similarity). Binds IGF2 more than IGF1. Forms a ternary complex of about 140 to 150 kDa with IGF1 or IGF2 and a 85 kDa glycoprotein (ALS). Interacts with HN. Interacts with TMEM219.

Family&Domains:

The thyroglobulin type-1 domain mediates interaction with HN.

Research Fields

· Cellular Processes > Cell growth and death > p53 signaling pathway.   (View pathway)

· Cellular Processes > Cell growth and death > Cellular senescence.   (View pathway)

· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.

References

1). Orai-IGFBP3 signaling complex regulates high-glucose exposureinduced increased proliferation, permeability, and migration of human coronary artery endothelial cells. BMJ Open Diabetes Research & Care, 2020 (PubMed: 33087338) [IF=4.1]

Application: WB    Species: Human    Sample: HCAECs

Figure 2 Orai1–3 protein and IGFBP3 expression levels and SOCE activity are significantly increased in HCAECs cultured in HG for 7 days and coronary artery endothelial cells of streptozotocin-induced diabetic mice with no change in STIM1 expression levels; increased Orais promotes proliferation of HCAECs and decreased IGFBP3 reduces proliferation of HCAECs. (A, C) Representative traces and (B, D) summary data showing the maximum increase in SOCE in HG-cultured HCAECs. After treatment of HCAECs with either 2 µM TG or 100 µM ATP for 10 min, application of 2 mM Ca2+ induces significant increase in [Ca2+]i through SOCE. (E, G, I, K, A') Representative western blot images and (F, H, J, L, B'–E') summary data showing Orai1, Orai2, Orai3, STIM1 and IGFBP3 expression levels in HCAECs cultured in NG or HG medium. (M, O, Q, F') Representative images of coronary artery endothelium immunostaining (brown; eg, red arrowheads) for protein expression levels of Orai1–3 and IGFBP3 in a mouse model of DM and control mice. Magnification, ×400. Scale bar, 50 µm, applies to all panels. (N, P, R, G') Quantification of Orai1–3 and IGFBP3 protein expression levels in the coronary artery endothelium of a mouse model of DM and control mice. (S) Representative western blot images and (T) summary data showing PCNA protein expression levels in HCAECs cultured in HG or NG medium for 7 days in the presence or absence of the SOCE agonist ATP (100 µM) or the SOCE inhibitor BTP2 (10 µM). (H') Representative western blot images and (I') summary data showing IGFBP3 siRNA knockdown of PCNA protein level. (U) Viability of HCAECs cultured in NG or HG medium for 7 days in the presence or absence of ATP (100 µM) or BTP2 (10 µM) as detected using the CCK-8 assay. (J') Results of CCK-8 assay examining HCAEC viability after cells were cultured in either NG or HG medium for 7 days and then transfected with either IGFBP3-specific siRNA or scrambled siRNA. β-tubulin or GAPDH was used as loading controls. Values represent mean±SEM (n=4–21 samples). *P<0.05, **P<0.01, ***P<0.001 compared with NG-cultured cells or control groups. BTP2, N-[4-[3,5-Bis(trifluoromethyl)-1H-pyrazol-1-yl]phenyl]-4-methyl-1,2,3-thiadiazole-5-carboxamide; CCK-8, Cell Count Kit-8; DM, diabetes mellitus; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; HCAECs, human coronary artery endothelial cells; HG, high glucose; IGFBP3, insulin-like growth factor binding protein 3; IOD, integrated optical density; NG, normal glucose; PCNA, proliferating cell nuclear antigen; siRNA, small interfering RNA; SOCE, store-operated Ca2+ entry; STIM1, stromal interaction molecule 1; TG, thapsigargin.

Application: IF/ICC    Species: Human    Sample: HCAECs

Figure 3 Physical interactions between the Orais and IGFBP3 proteins as examined by coimmunoprecipitation and immunofluorescence assays; SOCE regulates the expression level of IGFBP3 protein, which regulates the expression level of the Orai proteins in HCAECs cultured in HG medium. (A, C, E) HCAECs were cultured in HG medium for 7 days and then subjected to immunoprecipitation. Assays positive for Orai1, Orai2, or Orai3 proteins were used to pull down IGFBP3. (G, I, K) For reverse coimmunoprecipitation, immunoprecipitation assays positive for IGFBP3 were used to pull down Orai1, Orai2, or Orai3. (B, D, F, H, J, L) Summary data showing the statistical results of the corresponding coimmunoprecipitations. (M, O, Q) Representative confocal microscopy images showing co-localization analysis of anti-Orai1, 2, or 3 antibodies (red) and anti-IGFBP3 antibody (green). HCAECs were cultured in HG medium for 7 days and then fixed and incubated with the anti-Orai antibodies (red) and the anti-IGFBP3 antibody (green) and imaged using confocal microscopy. Representative confocal microscopy images of each antibody and the final merged images are shown. Nuclei are labeled blue with DAPI. (N, P, R) Fluorescence intensity profiles and summary data of anti-Orai antibodies (red) and the anti-IGFBP3 antibody (green) in the regions delineated by the corresponding yellow line. Scale bar, 10 mm. Co-localization area per cell was quantified using ImageJ. Representative western blot images (A') and summary data (B'–D') showing expression levels of the Orai proteins and SOCE activity induced by 100 µM ATP or 2 µM TG in HCAECs cultured in HG medium for 7 days after transfection with IGFBP3-specific siRNA or scrambled siRNA (siScrambled). Representative traces (E', G') and summary data (F', H') showing ATP-induced or TG-induced SOCE activity in HG-cultured HCAECs. Representative western blot images (I') and summary data (J') of IGFBP3 expression level in HCAECs cultured in HG medium for 7 days after the addition of the SOCE agonist ATP (100 µM) or the SOCE inhibitor BTP2 (10 µM). The same amount of solvent (ATP solvent was ddH2O (deionized distilled water); BTP2 solvent was DMSO (Dimethyl sulfoxide)) was used as control. (K') CCK-8 assay was used to detect the viability of HCAECs transfected with IGFBP3 siRNA (siIGFBP3) and cultured in medium with NG or HG for 7 days and treated with the SOCE agonist ATP (100 µM) or the SOCE inhibitor BTP2 (10 µM). Representative western blot images (L') and summary data (M') showing the expression levels of PCNA protein in HCAECs transfected with IGFBP3 siRNA and cultured in NG or HG medium for 7 days and then treated with ATP (100 µM) or BTP2 (10 µM). Values are mean±SEM (n=3–9 samples). *P<0.05, **P<0.01, ***P<0.001 compared with NG-cultured cells or control groups. BTP2, N-[4-[3,5-Bis(trifluoromethyl)-1H-pyrazol-1-yl]phenyl]-4-methyl-1,2,3-thiadiazole-5-carboxamide; CCK-8, Cell Count Kit-8; DAPI, 4',6-diamidino-2-phenylindole; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; HCAECs, human coronary artery endothelial cells; HG, high glucose; IGFBP3, insulin-like growth factor binding protein 3; NG, normal glucose; PCNA, proliferating cell nuclear antigen; siRNA, small interfering RNA; SOCE, store-operated Ca2+ entry; TG, thapsigargin.

2). Porcine IGFBP3 promotes porcine circovirus type 2 replication via PERK/eIF2α mediated DNA damage. Veterinary microbiology, 2023 (PubMed: 37922860) [IF=3.3]

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