VEGF antibody [VG-1] (ab1316)

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Anonymous Abreview submitted on 19 August 2008
Application:	Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
VEGF antibody [VG-1] (ab1316)
Sample	Human Tissue sections (Lung)
Specification	Lung
Fixative:	Paraformaldehyde
Antigen retrieval step:	Heat mediated - Buffer/Enzyme Used: Citrate buffer pH6
Permeabilization:	No
Blocking step:	Serum as blocking agent for  30 mins  · Concentration: 2 % · Temperature: 23°C
Primary Antibody
Dilution	1/100
Incubation time:	1 hour  Temperature: 23°C Diluent: PBS
Secondary Antibody
Name:	Non-Abcam Antibody was used: anti-mouse Conjugation: HRP
Additional Data
Rating	Good
Image	VEGF antibody [VG-1] for IHC-P (Human) Copyright image used under license

Anonymous Abreview submitted on 5 June 2008
Application:	Immunocytochemistry/ Immunofluorescence
VEGF antibody [VG-1] (ab1316)
Sample	Human Cell (MCF-7)
Specification	MCF-7
Fixative:	Paraformaldehyde
Permeabilization:	Yes - 0.1% Triton-X-100
Blocking step:	Serum as blocking agent for  1 hour  · Concentration: 2 %
Primary Antibody
Dilution	1/1000
Incubation time:	1 hour  Temperature: 37°C
Secondary Antibody
Name:	Non-Abcam Antibody was used: Alexa 546 conjugated goat anti-mouse IgG Conjugation: Alexa Fluor® 546
Dilution:	1/800
Additional Data
Rating	Excellent
Image	VEGF antibody [VG-1] for ICC/IF (Human) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 18 October 2007
Application:	Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
VEGF antibody [VG-1] (ab1316)
Sample	Rat Tissue sections (testes)
Specification	testes
Fixative:	Formaldehyde
Antigen retrieval step:	Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
Permeabilization:	No
Blocking step:	Serum as blocking agent for  30 mins  · Concentration: 1 % · Temperature: 22°C
Primary Antibody
Dilution	1/200
Incubation time:	30 minutes  Temperature: 22°C Diluent: PBS + 1% serum
Secondary Antibody
Name:	Non-Abcam Antibody was used: Goat anti-mouse HRP Conjugation: HRP
Dilution:	1/400
Additional Data
Rating	Good
Additional notes	Also worked well in cardiac muscle and kidney rat tissue (formulin fixed)
Image	VEGF antibody [VG-1] for IHC-P (Rat) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 18 October 2007
Application:	Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
VEGF antibody [VG-1] (ab1316)
Sample	Mouse Tissue sections (cystic kidney)
Specification	cystic kidney
Fixative:	Formaldehyde
Antigen retrieval step:	Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
Permeabilization:	No
Blocking step:	Serum as blocking agent for  30 mins  · Concentration: 1 % · Temperature: 22°C
Primary Antibody
Dilution	1/200
Incubation time:	30 minutes  Temperature: 22°C Diluent: PBS + 1% serum
Secondary Antibody
Name:	Non-Abcam Antibody was used: Goat anti-mouse HRP Conjugation: HRP
Dilution:	1/400
Additional Data
Rating	Average
Additional notes	As expected some background in mouse tissue. Would normally use a rabbit antibody for mouse IHC-P - however staining wasn't that bad under the circumstances.
Image	VEGF antibody [VG-1] for IHC-P (Mouse) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 15 January 2008
Application:	Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
VEGF antibody [VG-1] (ab1316)
Sample	Human Tissue sections (Adrenal tumour (VHL mutant))
Specification	Adrenal tumour (VHL mutant)
Fixative:	Formaldehyde
Antigen retrieval step:	Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
Permeabilization:	No
Blocking step:	Serum as blocking agent for  30 mins  · Concentration: 1 % · Temperature: 22°C
Primary Antibody
Dilution	1/200
Incubation time:	30 minutes  Temperature: 22°C Diluent: PBS + 1% serum
Secondary Antibody
Name:	Non-Abcam Antibody was used: Goat anti-rabbit HRP Conjugation: HRP
Dilution:	1/400
Additional Data
Rating	Excellent
Additional notes	We use subsequent peroxidase then protein block from Dako hence only the need for 1% serum - otherwise we find that 5% serum is easily sufficient as a block. In the case of this VEGF we normally use in situ hybridisation to look at mRNA to get cell specificity as VEGF is so readily secreted and most antibodies are 'dirty'. We tried this antibody with 5% serum and 1% + DAKO blocks and worked better with the latter.
Image	VEGF antibody [VG-1] for IHC-P (Human) Copyright image used under license

Anonymous Abreview submitted on 4 September 2007
Application:	Immunocytochemistry/ Immunofluorescence
VEGF antibody [VG-1] (ab1316)
Sample	Rat Cell (mesenchymal stem cell)
Specification	mesenchymal stem cell
Fixative:	Paraformaldehyde
Permeabilization:	Yes - Triton 0,3%
Blocking step:	Serum as blocking agent for  40 mins  · Concentration: 10 % · Temperature: 27°C
Primary Antibody
Dilution	1/1000
Incubation time:	2 hours  Temperature: 27°C Diluent: PBS
Secondary Antibody
Name:	Non-Abcam Antibody was used: goat anti-mouse IgG Conjugation: Cy3 ®
Dilution:	1/1000
Additional Data
Rating	Excellent
Additional notes	The secondary was a goat anti-mouse IgG, Cy3 in a 1:1000 dilution. And the block step was with normal goat serum.
Image	VEGF antibody [VG-1] for ICC/IF (Rat) Copyright image used under license

Anonymous Abreview submitted on 26 June 2007
Application:	Western blot
VEGF antibody [VG-1] (ab1316)
Sample	Human Cell lysate - whole cell (HL60)
Loading amount:	20 µg
Specification	HL60
Treatment	Arginase (6H)
Gel Running Conditions:	Reduced Denaturing
Blocking step:	Milk as blocking agent for  1 hour  · Concentration: 5 % · Temperature: RT°C
Primary Antibody
Dilution	1/1000
Incubation time:	16 hours  Temperature: 4°C Diluent: 5% milk TBS
Secondary Antibody
Name:	Non-Abcam Antibody was used: ECL Mouse IgG, HRP-Linked F(ab')2 Fragment Conjugation: HRP
Dilution:	1/2000
Detection
Detection method:	ECL
Exposure:	10 minutes and 0 seconds
Bands:	Specific: 51 kDa   Non-specific: 43, 60, 70, 80 kDa
Additional Data
Rating	Good
Additional notes	Cell lysed in RIPA buffer supplemented with Complete Protease Inhibitor Cocktail Tablets (Roche #11 836 153 001) & phosphatase inhibitors (Sigma #P2850 & P5726) Nitrocellulose membrane Some background. Could only clearly identify one of the cell associated isoforms (@ 51 kD). Nice signal though.
Image	VEGF antibody [VG-1] for WB (Human) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 7 February 2006
Application:	Immunoprecipitation
VEGF antibody [VG-1] (ab1316)
Sample	Dog Cell lysate - whole cell (kidney epithelium)
Specification	kidney epithelium
Immuno- precipitation step	Protein G
Primary Antibody
Dilution	1/1000
Incubation time:	2 hours
Western Blot Antibody
Name:	Non-Abcam Antibody was used: Goat anti-mouse IgG (HRP) Conjugation: HRP
Dilution:	1/1000
Additional Data
Rating	Good
Additional notes	The 50 kDa band was not visible using IP, perhaps a non-specific band in western blot only
Image	VEGF antibody [VG-1] for IP (Dog) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 7 February 2006
Application:	Immunoprecipitation
VEGF antibody [VG-1] (ab1316)
Sample	Rat Cell lysate - whole cell (dermal epithelium)
Specification	dermal epithelium
Immuno- precipitation step	Protein G
Primary Antibody
Dilution	1/1000
Incubation time:	2 hours
Western Blot Antibody
Name:	Non-Abcam Antibody was used: goat anti-mouse IgG (HRP) Conjugation: HRP
Dilution:	1/1000
Additional Data
Rating	Good
Image	VEGF antibody [VG-1] for IP (Rat) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 7 February 2006
Application:	Immunoprecipitation
VEGF antibody [VG-1] (ab1316)
Sample	Mouse Cell lysate - whole cell (primary retinal pigment epithelium)
Specification	primary retinal pigment epithelium
Immuno- precipitation step	Protein G
Primary Antibody
Dilution	1/1000
Incubation time:	2 hours
Western Blot Antibody
Name:	Non-Abcam Antibody was used: goat anti-mouse IgG (HRP) Conjugation: BSA
Dilution:	1/1000
Additional Data
Rating	Good
Additional notes	The higher band at 50 kDa was missing compared to western blots
Image	VEGF antibody [VG-1] for IP (Mouse) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 7 February 2006
Application:	Immunoprecipitation
VEGF antibody [VG-1] (ab1316)
Sample	Human Cell lysate - whole cell (retinal pigment epithelium line)
Specification	retinal pigment epithelium line
Immuno- precipitation step	Protein G
Primary Antibody
Dilution	1/1000
Incubation time:	2 hours
Western Blot Antibody
Name:	Non-Abcam Antibody was used: goat anti-mouse IgG (HRP) Conjugation: HRP
Dilution:	1/1000
Additional Data
Rating	Good
Additional notes	The 50 kDa band was missing using IP
Image	VEGF antibody [VG-1] for IP (Human) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 3 February 2006
Application:	Western blot
VEGF antibody [VG-1] (ab1316)
Sample	Rat Cell lysate - whole cell (dermal epithelium)
Specification	dermal epithelium
Blocking step:	Milk as blocking agent for  1 hour  · Concentration: 5%
Primary Antibody
Dilution	1/1000
Incubation time:	1 hour
Secondary Antibody
Name:	Non-Abcam Antibody was used: Goat anti-mouse IgG (HRP) Conjugation: HRP
Dilution:	1/1000
Detection
Detection method:	ECL
Exposure:	3 minutes and 0 seconds
Bands:	Specific: 28, 31, 50 kDa
Positive Control:	Primary rat dermal epithelial cells
Negative Control:	same cells, incubated without primary antibody
Additional Data
Rating	Good
Image	VEGF antibody [VG-1] for WB (Rat) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 3 February 2006
Application:	Western blot
VEGF antibody [VG-1] (ab1316)
Sample	Mouse Cell lysate - whole cell (retinal pigment epithelium)
Specification	retinal pigment epithelium
Blocking step:	Milk as blocking agent for  1 hour  · Concentration: 5%
Primary Antibody
Dilution	1/1000
Incubation time:	1 hour
Secondary Antibody
Name:	Non-Abcam Antibody was used: Goat anti-mouse IgG (HRP) Conjugation: HRP
Dilution:	1/1000
Detection
Detection method:	ECL
Exposure:	3 minutes and 0 seconds
Bands:	Specific: 28, 31, 50 kDa
Positive Control:	Primary mouse retinal pigment epithelium
Negative Control:	same cell type, incubated without primary antibody
Additional Data
Rating	Excellent
Image	VEGF antibody [VG-1] for WB (Mouse) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 6 February 2006
Application:	Western blot
VEGF antibody [VG-1] (ab1316)
Sample	Human Cell lysate - whole cell (retinal pigment epithelial line)
Specification	retinal pigment epithelial line
Blocking step:	Milk as blocking agent for  1 hour  · Concentration: 5%
Primary Antibody
Dilution	1/1000
Incubation time:	1 hour
Secondary Antibody
Name:	Non-Abcam Antibody was used: goat anti-mouse IgG (HRP) Conjugation: HRP
Dilution:	1/1000
Detection
Detection method:	ECL
Exposure:	3 minutes and 0 seconds
Bands:	Specific: 28, 31, 50 kDa
Positive Control:	Human retinal pigment epithelial cell line
Negative Control:	same cells, incubated without primary antibody
Additional Data
Rating	Excellent
Image	VEGF antibody [VG-1] for WB (Human) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 6 February 2006
Application:	Western blot
VEGF antibody [VG-1] (ab1316)
Sample	Dog Cell lysate - whole cell (canine kidney cell line)
Specification	canine kidney cell line
Blocking step:	Milk as blocking agent for  1 hour  · Concentration: 5%
Primary Antibody
Dilution	1/1000
Incubation time:	1 hour
Secondary Antibody
Name:	Non-Abcam Antibody was used: goat anti-mouse IgG (HRP) Conjugation: HRP
Dilution:	1/1000
Detection
Detection method:	ECL
Exposure:	3 minutes and 0 seconds
Bands:	Specific: 28, 31, 50 kDa
Positive Control:	Madin Darby canine kidney epithelial cell line lysate
Negative Control:	Same cell type, incubated without primary antibody
Additional Data
Rating	Excellent
Image	VEGF antibody [VG-1] for WB (Dog) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 21 November 2005
Application:	Immunocytochemistry
VEGF antibody [VG-1] (ab1316)
Sample	Dog Cell (kidney epithelium)
Specification	kidney epithelium
Fixative:	Paraformaldehyde
Blocking step:	Serum as blocking agent for  30 mins  · Concentration: 3%
Primary Antibody
Concentration	10 µg/ml
Incubation time:	3 hours
Secondary Antibody
Name:	Non-Abcam Antibody was used: Alexa Fluor 488 goat anti-mouse (IgG) Conjugation: Alexa Fluor® 488
Dilution:	1/200
Additional Data
Rating	Average
Image	VEGF antibody [VG-1] for ICC (Dog) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 18 November 2005
Application:	Immunocytochemistry
VEGF antibody [VG-1] (ab1316)
Sample	Cow Cell (retinal endothelium)
Specification	retinal endothelium
Fixative:	Paraformaldehyde
Blocking step:	Serum as blocking agent for  30 mins  · Concentration: 3%
Primary Antibody
Concentration	10 µg/ml
Incubation time:	2 hours
Secondary Antibody
Name:	Non-Abcam Antibody was used: Alexa Fluor 488 goat anti-mouse (IgG) Conjugation: Alexa Fluor® 488
Dilution:	1/200
Additional Data
Rating	Below Average
Additional notes	This antibody did not localize well to bovine VEGF. I would use another for cow cells.
Image	VEGF antibody [VG-1] for ICC (Cow) Copyright image used under license
Abcam response	Thank you for testing this antibody in different species; your other three reviews about this antibody have just been published. It seems that ab1316 works nicely in human, rat and mouse but it may not be suitable for detecting cow VEGF.

Anonymous (click to contact this user) Abreview submitted on 18 November 2005
Application:	Immunocytochemistry
VEGF antibody [VG-1] (ab1316)
Sample	Rat Cell (dermal epithelium)
Specification	dermal epithelium
Fixative:	Paraformaldehyde
Blocking step:	Serum as blocking agent for  30 mins  · Concentration: 3%
Primary Antibody
Concentration	10 µg/ml
Incubation time:	2 hours
Secondary Antibody
Name:	Non-Abcam Antibody was used: Alexa Fluor 488 goat anti-mouse (IgG) Conjugation: Alexa Fluor® 488
Dilution:	1/200
Additional Data
Rating	Good
Additional notes	This antibody showed distinct localization of VEGF on the membrane
Image	VEGF antibody [VG-1] for ICC (Rat) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 17 November 2005
Application:	Immunocytochemistry
VEGF antibody [VG-1] (ab1316)
Sample	Mouse Cell (retinal pigment epithelium)
Specification	retinal pigment epithelium
Fixative:	Paraformaldehyde
Blocking step:	Serum as blocking agent for  30 mins  · Concentration: 5%
Primary Antibody
Concentration	15 µg/ml
Incubation time:	3 hours
Secondary Antibody
Name:	Non-Abcam Antibody was used: Alexa Fluor 568 goat anti-mouse (IgG) Conjugation:
Dilution:	1/200
Additional Data
Rating	Average
Additional notes	wasn't as specific in mouse cells compared to human
Image	VEGF antibody [VG-1] for ICC (Mouse) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 16 November 2005
Application:	Immunocytochemistry
VEGF antibody [VG-1] (ab1316)
Sample	Human Cell (retinal pigment epithelium)
Specification	retinal pigment epithelium
Fixative:	Paraformaldehyde
Blocking step:	Serum as blocking agent for  30 mins  · Concentration: 3%
Primary Antibody
Concentration	10 µg/ml
Incubation time:	6 hours
Secondary Antibody
Name:	Non-Abcam Antibody was used: Alexa Fluor 568 goat anti-mouse (IgG) Conjugation:
Dilution:	1/200
Additional Data
Rating	Good
Image	VEGF antibody [VG-1] for ICC (Human) Copyright image used under license