Antibodies, Proteins, Kits and Reagents for Life Science

291 484 ₸ Product size

100 µl 291 484 ₸

Order now and get it on Tuesday March 02, 2021

Anti-Osteopontin antibody [EPR21139-316] (ab214050)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR21139-316] to Osteopontin
Suitable for: IP, IHC-P, WB
Reacts with: Human, Recombinant fragment

Product name

Anti-Osteopontin antibody [EPR21139-316]
See all Osteopontin primary antibodies
Description

Rabbit monoclonal [EPR21139-316] to Osteopontin
Host species

Rabbit

Tested Applications & Species

Application	Species
IHC-P	Human
IP	Human
WB	Human Recombinant fragment

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human Osteopontin recombinant protein (aa17-300) produced in a mammalian expression system; Human urine and fetal kidney lysates. IHC-P: Human kidney and liver tissues. IP: Human kidney lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR21139-316
Isotype

IgG
Research areas

Western blot - Anti-Osteopontin antibody [EPR21139-316] (ab214050)

All lanes : Anti-Osteopontin antibody [EPR21139-316] (ab214050) at 1/1000 dilution

Lane 1 : Human urine at 20 µg
Lane 2 : Human fetal kidney lysate at 10 µg

Secondary
Lane 1 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 35 kDa
Observed band size: 45,60,70,90 kDa
why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

The multiple bands observed are likely due to expression of different isoforms, as well as glycosylation (PMID: 8761944; PMID: 11375993; PMID: 8535240; PMID: 12890718).
Immunoprecipitation - Anti-Osteopontin antibody [EPR21139-316] (ab214050)

Osteopontin was immunoprecipitated from 0.35 mg of human kidney lysate with ab214050 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214050 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5,000 dilution

Lane 1: Human kidney lysate 10 µg (Input).

Lane 2: ab214050 IP in human kidney lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214050 in human kidney lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 minutes.

The multiple bands observed are likely due to expression of different isoforms, as well as glycosylation (PMID: 8761944; PMID: 11375993; PMID: 8535240; PMID: 12890718).
Western blot - Anti-Osteopontin antibody [EPR21139-316] (ab214050)

Anti-Osteopontin antibody [EPR21139-316] (ab214050) at 1/1000 dilution + Human Osteopontin recombinant protein (aa17-300) produced in a mammalian expression at 0.01 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 35 kDa
Observed band size: 60-65 kDa why is the actual band size different from the predicted?

Exposure time: 26 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The his-tagged recombinant protein was produced in a mammalian expression system, thereby allowing it to be post-translationally modified.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21139-316] (ab214050)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Osteopontin with ab214050 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in bile ducts of human liver (PMID: 16221502). Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody [EPR21139-316] (ab214050)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Osteopontin with ab214050 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use. Cytoplasmic staining in distal tubules of human kidney (PMID: 17938278), performed on a Leica Biosystems BOND® RX instrument. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat-mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 minutes.
Anti-Osteopontin antibody [EPR21139-316] (ab214050)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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