Osteopontin antibody (ab8448)

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Anonymous (click to contact this user) Abreview submitted on 5 November 2009
Application:	Immunohistochemistry (Frozen sections)
Osteopontin antibody (ab8448)
Sample	Mouse Tissue sections (Developing skeleton)
Specification	Developing skeleton
Fixative:	Paraformaldehyde
Permeabilization:	Yes - 0.1% Triton
Blocking step:	Serum as blocking agent for  1 hour  · Concentration: 20 % · Temperature: RT°C
Primary Antibody
Dilution	1/200
Incubation time:	16 hours  Temperature: 4°C Diluent: 20%FBS/PBS
Secondary Antibody
Name:	Non-Abcam Antibody was used: Alexa Fluoro 488 Conjugation: Alexa Fluor® 488
Dilution:	1/200
Additional Data
Rating	Excellent
Additional notes	Image: Red Rhodamine Phalloidin (muscle), Blue DAPI (nuclei), Green Osteopontin
Image	Osteopontin antibody for IHC-Fr (Mouse) Copyright image used under license

review by Judith Kranz submitted on 18 February 2009
Application:	Immunohistochemistry (Frozen sections)
Osteopontin antibody (ab8448)
Sample	Rat Tissue sections (Kidney)
Specification	Kidney
Fixative:	Paraformaldehyde
Permeabilization:	No
Blocking step:	Serum as blocking agent for  1 hour  · Concentration: 10 % · Temperature: 23°C
Primary Antibody
Dilution	1/200
Incubation time:	16 hours  Temperature: 4°C Diluent: TBS
Secondary Antibody
Name:	Non-Abcam Antibody was used: Jackson TRITC-conjugated Donkey anti-Rabbit Conjugation: TRITC
Dilution:	1/200
Additional Data
Rating	Good
Additional notes	Rat 4% PFA-fixed frozen OR paraffin sections have worked successfully with a 1/200 dilution of this antibody. Image shows strong OPN staining of some renal tubules, and some accumulation of OPN in the glomerulus. DAPI-stained nuclei are blue.
Image	Osteopontin antibody for IHC-Fr (Rat) Copyright image used under license

review by Werner Goetz submitted on 5 September 2008
Application:	Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Osteopontin antibody (ab8448)
Sample	Rat Tissue sections (jaw bone (mandible) with teeth)
Specification	jaw bone (mandible) with teeth
Fixative:	Paraformaldehyde
Antigen retrieval step:	Enzymatic - Buffer/Enzyme Used: 0.4% pepsin 37°C
Permeabilization:	No
Blocking step:	BSA as blocking agent for  20 mins  · Concentration: 4 % · Temperature: rt°C
Primary Antibody
Dilution	1/100
Incubation time:	1 hour  Diluent: TBS/BSA
Secondary Antibody
Name:	Non-Abcam Antibody was used: EnVision Conjugation: HRP
Additional Data
Rating	Good
PubMed ID	18632841: View
Image	Osteopontin antibody for IHC-P (Rat) Copyright image used under license

Anonymous Abreview submitted on 31 January 2007
Application:	Western blot
Osteopontin antibody (ab8448)
Sample	Rat Tissue lysate - whole (skin)
Loading amount:	15 µg
Specification	skin
Blocking step:	Milk as blocking agent for  1 hour   30 mins  · Concentration: 5%
Primary Antibody
Dilution	1/1000
Incubation time:	12 hours
Secondary Antibody
Name:	Non-Abcam Antibody was used: Monoclonal mouse anti rabbit Conjugation: Alkaline Phosphatase
Dilution:	1/1000
Detection
Detection method:	npt/bcip
Exposure:	30 minutes and 0 seconds
Bands:	Specific: 30 kDa
Positive Control:	Ran 4 varoius samples (2 of each kind) and observed band on 2 of the samples of the same kind.
Additional Data
Rating	Average
Additional notes	Antibody buffer for primary and secondary was 1.5% milk in TBS-T. Used 10% gel and a PVDF Membrane was use as blot paper.
Image	Osteopontin antibody for WB (Rat) Copyright image used under license

Anonymous Abreview submitted on 16 February 2007
Application:	Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Osteopontin antibody (ab8448)
Sample	Human Tissue sections (Breast)
Specification	Breast
Fixative:	Formaldehyde
Antigen retrieval step:	Heat mediated
Blocking step:	Serum as blocking agent for  45 mins  · Concentration: 10%
Primary Antibody
Dilution	1/750
Incubation time:	1 hour
Secondary Antibody
Name:	Non-Abcam Antibody was used: From Vectastain Elite kit anti-rabbit Conjugation: HRP polymer
Dilution:	1/200
Additional Data
Rating	Good
Additional notes	Endogenous peroxidase activity was blocked with 3% hydrogen peroxide in methanol for 5 min.
Image	Osteopontin antibody for IHC-P (Human) Copyright image used under license

review by Melinda Merrell submitted on 29 November 2004
Application:	Western blot
Osteopontin antibody (ab8448)
Additional Data
Notes	Overall a very good antibody for Western blotting. Some non-specific binding, but the correct bands, as indicated by the product sheet, were readily found. Very good detection - sharp, clear bands with very little exposure to film time. Only slight problem detected would be an apparent interaction with bovine serum...in a few samples I had made in serum, there were very large "blobs" of protein around 66kD, which we assumed to be an interaction with the bovine serum (due to the fact that in the serum-starved samples, we did not see the same thing). That would be the only potential drawback to this problem.
Dilution notes	1:1000 in 1% BSA/TBST

review by Bart Tambuyzer submitted on 25 June 2004
Application:	Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Osteopontin antibody (ab8448)
Additional Data
Notes	This antibody readily stained a cell population within the porcine neonatal brain. These cells - however we are still investigating them - are presumably microglia. The best stainings were observed with formalin fixed or Bouin fixed paraffin embedded sections of the brain. We also detected OPN in supernatant of LLC-PK1 cells (porcine renal celline) using the dotblot technique.
Dilution notes	Immunohistochemistry: 1/4000 Dotblot: 1/2000