Home > Product > Antibody > Rabbit Anti-Osteopontin antibody
BNSP; Bone sialoprotein 1; BSP I; BSPI; Early T lymphocyte activation 1; ETA 1; ETA1; MGC110940; Nephropontin; OPN; osteopontin/immunoglobulin alpha 1 heavy chain constant region fusion protein; OSTP_HUMAN; secreted phosphoprotein 1 (osteopontin, bone sia
Cat:
SL0026R
Species Reactivity:
Human,Mouse,Rat,
Immunogen:
KLH conjugated synthetic peptide derived from mouse OPN:221-280/294
Format:
Liquid
Storage instructions:
Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
Concentration:
1mg/ml
Clonality:
Polyclonal
Isotype:
IgG
Applications:
WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500IHC-F=1:100-500Flow-Cyt=2ug/TestIF=1:100-500(Paraffin sections need to do antigen repair)not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.
Host:
Rabbit
Product Overview:
Sample: Lane 1: HepG2 (Human) Cell Lysate at 30 ugLane 2: A549 (Human) Cell Lysate at 30 ugLane 3: U251 (Human) Cell Lysate at 30 ugPrimary: Anti-Osteopontin (SL0026R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 34 kDObserved band size: 60 kDParaformaldehyde-fixed, paraffin embedded (Rat lung); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Osteopontin) Polyclonal Antibody, Unconjugated (SL0026R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.Tissue/cell: rat shin bone; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti-osteopontin Polyclonal Antibody, Unconjugated(SL0026R) 1:300, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) stainingTissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti-osteopontin Polyclonal Antibody, Unconjugated(SL0026R) 1:300, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) stainingBlank control: HepG2. Primary Antibody (green line): Rabbit Anti-Osteopontin antibody (SL0026R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG .Secondary Antibody : Goat anti-rabbit IgG-AF647Dilution: 1μg /test. ProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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Product PDFs
Datasheet:


Osteopontin is the principal phosphorylated glycoprotein of bone and is expressed in a limited number of other tissues including dentine. Osteopontin is produced by osteoblasts under stimulation by calcitriol and binds tightly to hydroxyapatite. It is also involved in the anchoring of osteoclasts to the mineral of bone matrix via the vitronectin receptor, which has specificity for osteopontin. Osteopontin is overexpressed in a variety of cancers, including lung, breast, colorectal, stomach, ovarian, melanoma and mesothelioma.

Subunit:
Ligand for integrin alpha-V/beta-3.

Subcellular Location:
Secreted.

Tissue Specificity:
Bone. Found in plasma.

Similarity:
Belongs to the osteopontin family.

SWISS:
P10923

Gene ID:
20750

Database links:

Entrez Gene: 6696 Human

Entrez Gene: 20750 Mouse

Entrez Gene: 25353 Rat

Omim: 166490 Human

SwissProt: P10451 Human

SwissProt: P10923 Mouse

SwissProt: P08721 Rat

Unigene: 313 Human



骨桥蛋白(osteopontin)又称为早期T淋巴细胞活化因子I或分泌型磷蛋白I,是一种重要的多功能细胞外基质蛋白,在骨组织细胞、巨噬细胞、平滑肌细胞、内皮细胞及上皮细胞等多种细胞上表达,在介导细胞的趋化、聚集、黏附、增殖和迁移,以及骨组织的矿化与重建、免疫调节、信号转导和对感染性疾病的免疫能力等方面有着重要的作用。
OPN在细胞免疫中的功能:1.OPN对炎性细胞的作用:炎症是炎性细胞(单核巨噬细胞、淋巴细胞、中性粒细胞等)在炎症部位的聚集,适度的炎症对机体有益,如抗感染及促进组织细胞功能恢复等;但过度的炎症能引起组织损伤等有害作用。OPN在炎症中具有促炎及抗炎双重作用;2.OPN对淋巴细胞的作用:早期的报道称OPN是T淋巴细胞抑制因子,它可抑制T淋巴细胞而促进B淋巴细胞及肥大细胞的分化、增殖
骨桥蛋白(osteopontin,OPN)是一种含精氨酸—甘氨酸—天冬氨酸(Arg-Gly-Asp,RGD)的分泌型糖基化磷蛋白,已归类于细胞外基质(extracellular matrix,ECM)蛋白。 OPN既是细胞因子,又是细胞黏附分子。
OPN最初是从骨组织分离的,而后在上皮细胞、胃肠道的分泌物、肾、甲状腺、睾丸以及一些肿瘤细胞里都陆续的发现了OPN的表达。在生殖方面,OPN在分泌期子宫内膜上皮细胞、侵袭的滋养细胞、蜕膜细胞中表达。 Picture

Sample:
Lane 1: HepG2 (Human) Cell Lysate at 30 ug
Lane 2: A549 (Human) Cell Lysate at 30 ug
Lane 3: U251 (Human) Cell Lysate at 30 ug
Primary: Anti-Osteopontin (SL0026R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 34 kD
Observed band size: 60 kD
Paraformaldehyde-fixed, paraffin embedded (Rat lung); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Osteopontin) Polyclonal Antibody, Unconjugated (SL0026R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Tissue/cell: rat shin bone; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-osteopontin Polyclonal Antibody, Unconjugated(SL0026R) 1:300, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining
Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-osteopontin Polyclonal Antibody, Unconjugated(SL0026R) 1:300, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining
Blank control: HepG2. Primary Antibody (green line): Rabbit Anti-Osteopontin antibody (SL0026R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-AF647 Dilution: 1μg /test. Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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