bs-0019R [Primary Antibody]
Rabbit  Anti-Osteopontin Rabbit pAb  Polyclonal Antibody
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Host: Rabbit

Target Protein: Osteopontin Rabbit pAb

IR: Immunogen Range:141-220/314

Clonality: Polyclonal

Isotype: IgG

Entrez Gene: 6696

Swiss Prot: P10451

Source: KLH conjugated synthetic peptide derived from human OPN:141-220/314 

Purification: affinity purified by Protein A

Storage: 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles.

Background: 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.

Size: 100ul

Concentration: 1mg/ml

Applications: WB=1:500-2000,IHC-P=1:500-2000,IHC-F=1:100-500,IF=1:100-500,Flow-Cyt=1ug/Test

Cross Reactive Species: Human,Rat

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES
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 (bs-0052R) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 60-65 kD Observed band size: 60 kD
Sample: A549 Cell Lysate at 40 ug Primary: Anti-ostepontin (bs-0019R) at 1/300 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 35 kD Observed band size: 50 kD
Paraformaldehyde-fixed, paraffin embedded Rat lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Osteopontin Polyclonal Antibody, Unconjugated (bs-0019R) at 1:400 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Blank control: HepG2. Primary Antibody (green line): Rabbit Anti-Osteopontin antibody (bs-0019R) 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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