Osteopontin/OPN Antibody (1B20) - BSA Free

Novus Biologicals | Catalog # NB110-89062

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Rabbit

Cited:

Human, Mouse, Rat, Bovine, Rabbit

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 1B20

Format

BSA Free
View all Osteopontin/OPN Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide corresponding to the C-terminus of human Osteopontin [Swiss-Prot# P10451].

Reactivity Notes

Human, Rat and Rabbit. Rabbit reactivity reported in scientific literature (PMID:33001866). Use in Mouse reported in scientific literature (PMID:33580126).

Localization

Secreted.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Osteopontin/OPN Antibody (1B20) - BSA Free

Western Blot: Osteopontin/OPN Antibody (1B20)BSA Free [NB110-89062]

Western Blot: Osteopontin/OPN Antibody (1B20)BSA Free [NB110-89062]

Western Blot: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062] - Analysis of Osteopontin expression in U2OS whole cell lysate.
Immunocytochemistry/ Immunofluorescence: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062]

Immunocytochemistry/ Immunofluorescence: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062]

Immunocytochemistry/Immunofluorescence: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062] - Osteopontin antibody was tested at 1:50 in U2OS cells with FITC (green). Nuclei and actin were counterstained with Dapi (blue) and Phalloidin (red).
Immunohistochemistry: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062]

Immunohistochemistry: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062]

Osteopontin-OPN-Antibody-1B20-BSA-Free-Immunohistochemistry-NB110-89062-img0012.jpg
Immunohistochemistry: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062]

Immunohistochemistry: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062]

Osteopontin-OPN-Antibody-1B20-BSA-Free-Immunohistochemistry-NB110-89062-img0013.jpg
Immunohistochemistry: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062]

Immunohistochemistry: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062]

Immunohistochemistry: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062] - Analysis of Osteopontin on human lung adenocarcinoma.
Immunohistochemistry: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062]

Immunohistochemistry: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062]

Osteopontin-OPN-Antibody-1B20-BSA-Free-Immunohistochemistry-NB110-89062-img0011.jpg
Simple Western: Osteopontin/OPN Antibody (1B20)BSA Free [NB110-89062]

Simple Western: Osteopontin/OPN Antibody (1B20)BSA Free [NB110-89062]

Simple Western: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062] - Simple Western lane view shows a specific band for Osteopontin in 0.5 mg/ml of Human Breast Cancer lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system. * Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody
Osteopontin/OPN Antibody (1B20) - BSA Free

Immunohistochemistry: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062] -

Immunohistochemistry: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062] - Photomicrographs of immunostaining for osteocalcin & osteopontin in (A) Group I, (B) Group II (C) Group III at 6 weeks.The pictures are arranged by staining technique (columns) & the investigated treatment (rows). Areas that stained positive for osteocalcin & osteopontin are indicated by red arrowheads. NB, new bone; OS, osteoid; FM, fatty marrow. Image collected & cropped by CiteAb from the following publication (https://peerj.com/articles/3513), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Osteopontin/OPN Antibody (1B20) - BSA Free

Immunohistochemistry: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062] -

Immunohistochemistry: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062] - Photomicrographs of immunostaining for osteocalcin & osteopontin in (A) Group I, (B) Group II (C) Group III at 6 weeks.The pictures are arranged by staining technique (columns) & the investigated treatment (rows). Areas that stained positive for osteocalcin & osteopontin are indicated by red arrowheads. NB, new bone; OS, osteoid; FM, fatty marrow. Image collected & cropped by CiteAb from the following publication (https://peerj.com/articles/3513), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Osteopontin/OPN Antibody (1B20) - BSA Free

Western Blot: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062] -

Western Blot: Osteopontin/OPN Antibody (1B20) - BSA Free [NB110-89062] - Photomicrographs of immunostaining for osteocalcin & osteopontin in (A) Group I, (B) Group II (C) Group III at 6 weeks.The pictures are arranged by staining technique (columns) & the investigated treatment (rows). Areas that stained positive for osteocalcin & osteopontin are indicated by red arrowheads. NB, new bone; OS, osteoid; FM, fatty marrow. Image collected & cropped by CiteAb from the following publication (https://peerj.com/articles/3513), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Osteopontin/OPN Antibody (1B20) - BSA Free

Osteopontin/OPN (1B20) in HepG2 Human Cell Line.

Osteopontin/OPN (1B20) was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Rat anti- Osteopontin/OPN (1B20) Protein G Purified Monoclonal Antibody conjugated to Alexa Fluor® 488 (Catalog # NB110-89062AF488) (green) at 10 µg/mL overnight at 4C. Cells were counterstained with DAPI (dark blue). Cells were imaged using a 100X objective and digitally deconvolved.

Applications for Osteopontin/OPN Antibody (1B20) - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:50

Immunohistochemistry

1:100

Immunohistochemistry-Paraffin

1:100

Immunoprecipitation

1:10-1:500

Simple Western

1:20

Western Blot

1:1000

Reviewed Applications

Read 2 reviews rated 5 using NB110-89062 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Osteopontin/OPN

The matricellular protein osteopontin binds to cell surface receptors and is secreted into many body fluids including milk, blood, and urine, depending on the organ of origin. This makes osteopontin an ideal candidate for being a biomarker as the secreted form is easily obtained in throwaway fluids, and mimics the cellular environment from which it is released. Osteopontin is important in immune responses and inflammation as well as bone generation and remodeling. In autistic children, serum levels of osteopontin are correlated to the severity of disease, probably due to a brain inflammation pattern in these children. In aortic valve sclerosis and stenosis, increased levels of secreted osteopontin are also noted. Osteopontin has also been suggested as a cancer biomarker, since it is associated with tumor formation, progression and metastasis. In bone and tooth formation osteopontin is known to be a negative regulator of parathyroid hormone-related protein receptor, which induces osteogenesis. Without appropriate levels of osteopontin, bone growth continues unregulated, and leads to specific bone cancers. In short, osteopontin is a strong marker for bone growth, inflammation and certain cancers.

Long Name

Secreted Phosphoprotein 1 [BNSP]

Alternate Names

Eta-1, OPN, Spp1

Entrez Gene IDs

6696 (Human); 25353 (Rat)

Gene Symbol

SPP1

UniProt

P08721, p10451 (Human)

Additional Osteopontin/OPN Products

Product Documents for Osteopontin/OPN Antibody (1B20) - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for Osteopontin/OPN Antibody (1B20) - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for Osteopontin/OPN Antibody (1B20) - BSA Free

Customer Reviews for Osteopontin/OPN Antibody (1B20) - BSA Free (2)

5 out of 5
2 Customer Ratings
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Customer Images

Showing  1 - 2 of 2 reviews Showing All
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  • Osteopontin/OPN Antibody (1B20)
    Name: Anonymous
    Application: Immunofluorescence - fixed-frozen
    Sample Tested: bone marrow
    Species: Mouse
    Verified Customer | Posted 02/29/2020
    Mouse OPN 1:100
    Mouse bones were fixed in PFA 4% for 4h on ice, decalcified in EDTA for 24h at 4°C, cryoprotected in 30% sucrose for 24 at 4°C, and embedded in OCT
    Osteopontin/OPN Antibody (1B20) - BSA Free NB110-89062
  • Osteopontin/OPN Antibody (1B20)
    Name: Cindy Zhou
    Application: ELISA
    Sample Tested: Human breast carcinoma and Human breast cancer plasma
    Species: Human
    Verified Customer | Posted 08/11/2017
    It will be nice if using PBS buffer for product. Removing Gly-Tris buffer is a pain.

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Protocols

View specific protocols for Osteopontin/OPN Antibody (1B20) - BSA Free (NB110-89062):


Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.


Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures."


Western Blot Protocol
1. Perform SDS-PAGE on samples to be analyzed, loading 30 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.


Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Osteopontin/OPN Antibody (1B20) - BSA Free

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  • Q: I am very interested in your anti-human osteopontin (SPP1) (cat #NB110-89062). I work with bovine and would like to use the antibody in the blot. According to your description, the immunogen is a synthetic peptide which binds to the C-terminus of the human. The sequences of the C-termini are very similar in humans and cattle. Could you kindly tell me what amino acids against the C-terminus of the human antibody is directed exactly?

    A:

    The immunogen sequence for NB110-89062 lies within amino acids 284-314 of isoform A, UniProt P10451. I have carried out a UniProt Blast analysis of the exact immunogen sequence versus Bos taurus (bovine) proteins for you, and the result was a 95% match for both osteopontin and osteopontin-K. This is a high percentage strongly suggesting that the antibody will be suitable for bovine, although we cannot guarantee it as this has not been tested in the lab. Because you would be using NB110-89062 with a species not stated on the web page / datasheet for this product, you would not be covered by the Novus Guarantee but you would be eligible for our special Innovator's Reward program, where as a thank you for sharing your results with us (whether the antibody proved suitable or not) by completing a review, we would give you a voucher code for the price of the product. Please contact us at innovators@novusbio.com for more information.

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