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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Flow cytometry [1]
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- Product number
- 702184 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- OPN-R Recombinant Rabbit Monoclonal Antibody (24H5L3)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with Monkey, Rabbit and Bat.
- Antibody clone number
- 24H5L3
- Concentration
- 0.5 mg/mL
Submitted references PLK2 modulation of enriched TAp73 affects osteogenic differentiation and prognosis in human osteosarcoma.
Li W, Zhang X, Xi X, Li Y, Quan H, Liu S, Wu L, Wu P, Lan W, Shao Y, Li H, Chen K, Hu Z
Cancer medicine 2020 Jun;9(12):4371-4385
Cancer medicine 2020 Jun;9(12):4371-4385
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of U-2-OS (Lane 1), Hep G2 (Lane 2), MKN45 (Lane 3) and Mouse stomach (Lane 4). The blots were probed with Anti-Osteopontin Recombinant Rabbit Monoclonal Antibody (Product # 702184, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 35 kDa band corresponding to Osteopontin was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis was performed on fixed and permeabilized U-2 OS cells for detection of endogenous osteopontin (cleaved) using Anti-osteopontin (cleaved) Recombinant Rabbit Monoclonal Antibody (Product # 702184, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of cleaved osteopontin protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of cleaved osteopontin. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometry analysis of endogenous Osteopontin (cleaved) was performed on U-2 OS cells labeled with ABfinity™ Anti-Osteopontin (cleaved) Recombinant Rabbit Monoclonal Antibody (Product# 702183, 5 ug/ 1M cells) or with rabbit isotype control at 0.5 ug/ml and detected with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, (Alexa Fluor® 488 conjugate, Product# A27034, 0.4 ug/ml, 1:2500) as represented by the red and pink histograms respectively. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control. A representative of 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer (4468770).
