Anti-Osteopontin antibody (ab8448)
Key features and details
- Rabbit polyclonal to Osteopontin
- Suitable for: IHC-P, ICC/IF, WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Osteopontin antibody
See all Osteopontin primary antibodies -
Description
Rabbit polyclonal to Osteopontin -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIHC-P HumanWB Human
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Immunogen
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
Preservative: 0.1% Sodium azide
Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride -
Concentration information loading... -
Purity
Whole antiserum -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Osteopontin antibody (ab8448)
Human Breast tumour section. Osteopontin is a normal component of elastic fibers in the breast (shown heavily stained in this section). There is also weak staining of the extracellular matrix. Osteopontin is not believed to be expressed inside breast tumour cells, and there is no staining in the intracellular region of the breast cells in this section.
Osteopontin antibody (ab8448) used at 1:100-1:300. No antigen retrieval is required.
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Western blot - Anti-Osteopontin antibody (ab8448)All lanes : Anti-Osteopontin antibody (ab8448) at 1/1000 dilution
Lane 2 : Human Osteopontin
Lane 3 : MMP-cleaved Human Osteopontin
Lysates/proteins at 0.25 µg per lane.
Secondary
All lanes : HRP-conjugated Goat anti-Rabbit IgG at 1/10000 dilution
The osteopontin antibody (ab8448) is used at 1:1000 dilution on a blot with 250ng human osteopontin (lane 2)and MMP-cleaved osteopontin (lane 3 -
Immunocytochemistry/ Immunofluorescence - Anti-Osteopontin antibody (ab8448)ICC/IF image of ab8448 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab8448, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Cellular activity - Anti-Osteopontin antibody (ab8448) This image is courtesy of Agnihotri et al, JBC 2001OPN is cleaved by MMP to yield 2 fragments, which migrate at 40kD(N terminal) and 32kD (C terminal). The C terminal fragment can undergo further cleavage by both of these MMPs (see Agnihotri et al, JBC 2001 for further details). The epitope recognised by ab8448 is shown in violet. This antibody detects the full length OPN and the 32kD fragment. It does not recognise the 40kD fragment.
