Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker (ab2733)

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Anonymous (click to contact this user) Abreview submitted on 4 February 2010
Application:	Immunocytochemistry/ Immunofluorescence
Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker (ab2733)
Sample	Human Cell (HEK293 cell)
Specification	HEK293 cell
Fixative:	Paraformaldehyde
Permeabilization:	Yes - 0.5% Triton X-100
Blocking step:	Serum as blocking agent for  1 hour  · Concentration: 5 % · Temperature: 24°C
Primary Antibody
Dilution	1/500
Incubation time:	1 hour  Temperature: 37°C Diluent: 5% horse serum 0.1% NP-40 in PBS
Secondary Antibody
Name:	Non-Abcam Antibody was used: goat-anti-mouse Alexa Fluor 555 Conjugation: Alexa Fluor® 555
Dilution:	1/1000
Additional Data
Rating	Excellent
Additional notes	This antibody is great to detect endosomal structures in HEK293 cells
Image	Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker for ICC/IF (Human) Copyright image used under license

Anonymous Abreview submitted on 21 May 2009
Application:	Immunocytochemistry/ Immunofluorescence
Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker (ab2733)
Sample	Mouse Cell (blastocyst)
Specification	blastocyst
Fixative:	Formaldehyde
Permeabilization:	Yes - tritonX-100
Blocking step:	BSA as blocking agent for  1 hour  · Concentration: 2 % · Temperature: 4°C
Primary Antibody
Dilution	1/100
Incubation time:	1 hour  Temperature: 4°C
Secondary Antibody
Name:	Non-Abcam Antibody was used: Alexa Fluor 488 goat anti-mouse Conjugation: Alexa Fluor® 488
Dilution:	1/250
Additional Data
Rating	Good
Image	Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker for ICC/IF (Mouse) Copyright image used under license

Anonymous Abreview submitted on 1 December 2008
Application:	Immunocytochemistry/ Immunofluorescence
Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker (ab2733)
Sample	Rat Cell (Rat2 Cells)
Specification	Rat2 Cells
Fixative:	Paraformaldehyde
Permeabilization:	Yes - 0.1% Saponin
Blocking step:	BSA as blocking agent for  1 hour  · Concentration: 0.1 % · Temperature: 37°C
Primary Antibody
Dilution	1/500
Incubation time:	1 hour  Temperature: 37°C Diluent: PBS + 0.1% BSA
Secondary Antibody
Name:	Non-Abcam Antibody was used: alexa 555 donkey anti mouse Conjugation: Alexa Fluor® 555
Dilution:	1/1000
Additional Data
Rating	Excellent
Additional notes	Excellent, specific staining.
Image	Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker for ICC/IF (Rat) Copyright image used under license

Anonymous (click to contact this user) Abreview submitted on 12 December 2008
Application:	Immunocytochemistry/ Immunofluorescence
Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker (ab2733)
Sample	Rat Cell (Rat2 Cells)
Specification	Rat2 Cells
Fixative:	Paraformaldehyde
Permeabilization:	Yes - 0.1% Triton
Blocking step:	BSA as blocking agent for  16 hours  · Concentration: 0.1 % · Temperature: 4°C
Primary Antibody
Dilution	1/500
Incubation time:	1 hour  Temperature: 37°C Diluent: PBS + 0.1% BSA
Secondary Antibody
Name:	Non-Abcam Antibody was used: Alexa Fluor 488 donkey anti-mouse Conjugation: Alexa Fluor® 488
Dilution:	1/1000
Additional Data
Rating	Good
Additional notes	Endosomes are in green and nucleus is in blue (DAPI).
Image	Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker for ICC/IF (Rat) Copyright image used under license

Anonymous Abreview submitted on 10 October 2008
Application:	Immunocytochemistry/ Immunofluorescence
Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker (ab2733)
Sample	Human Cell (MDA-MB-231 breast cancer cell line)
Specification	MDA-MB-231 breast cancer cell line
Fixative:	Paraformaldehyde
Permeabilization:	Yes - Triton 0.03%
Blocking step:	Serum as blocking agent for  45 mins  · Concentration: 10 % · Temperature: 25°C
Primary Antibody
Dilution	1/200
Incubation time:	45 minutes  Temperature: 25°C Diluent: PBS+10%FBS
Secondary Antibody
Name:	Non-Abcam Antibody was used: goat-anti-mouse-Alexa488 Conjugation: Alexa Fluor® 488
Dilution:	1/1500
Additional Data
Rating	Excellent
Image	Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker for ICC/IF (Human) Copyright image used under license

Anonymous Abreview submitted on 18 April 2006
Application:	Immunocytochemistry/ Immunofluorescence
Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker (ab2733)
Sample	Dog Cell (MDCK (epithelial kidney))
Specification	MDCK (epithelial kidney)
Fixative:	Paraformaldehyde
Blocking step:	BSA as blocking agent for  1 hour  · Concentration: 3%
Primary Antibody
Concentration	2 µg/ml
Incubation time:	3 hours
Secondary Antibody
Name:	Non-Abcam Antibody was used: Alexa Fluor 488 Conjugation: Alexa Fluor® 488
Dilution:	1/500
Additional Data
Rating	Average
Additional notes	this antibody is not great for MDCK cells but it works decent
Image	Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker for ICC/IF (Dog) Copyright image used under license

Anonymous Abreview submitted on 18 April 2006
Application:	Immunocytochemistry/ Immunofluorescence
Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker (ab2733)
Sample	Human Cell (Hela)
Specification	Hela
Fixative:	Paraformaldehyde
Blocking step:	BSA as blocking agent for  1 hour  · Concentration: 3
Primary Antibody
Concentration	2 µg/ml
Incubation time:	3 hours
Secondary Antibody
Name:	Non-Abcam Antibody was used: AlexaFluor488 Conjugation: Alexa Fluor® 488
Dilution:	1/500
Additional Data
Rating	Good
Image	Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker for ICC/IF (Human) Copyright image used under license

review by Kun Liu submitted on 19 September 2005
Application:	Immunocytochemistry
Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker (ab2733)
Sample	Human Cell (HEK 293 Cell line)
Specification	HEK 293 Cell line
Fixative:	Formaldehyde
Blocking step:	Serum as blocking agent for  1 hour  · Concentration: 2%
Primary Antibody
Dilution	1/200
Incubation time:	1 hour
Secondary Antibody
Name:	Non-Abcam Antibody was used: Alexa 546 conjugated IgG Conjugation:
Dilution:	1/1500
Additional Data
Rating	Excellent
Additional notes	2%BSA plus 2% goat serum as the blocking solution.
Image	Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker for ICC (Human) Copyright image used under license

review by Kun Liu submitted on 23 August 2005
Application:	Immunocytochemistry
Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker (ab2733)
Sample	Dog Cell lysate - whole cell (MDCK cell line)
Specification	MDCK cell line
Fixative:	Formaldehyde
Blocking step:	Serum as blocking agent for  1 hour  · Concentration: 2%
Primary Antibody
Dilution	1/200
Incubation time:	1 hour
Secondary Antibody
Name:	Non-Abcam Antibody was used: Alexa Fluor 546 goat anti mouse IgG (H+L) Conjugation:
Dilution:	1/2000
Additional Data
Rating	Excellent
Additional notes	I used this antibody for immunofluorecence in MDCK cell line. Actually I used 2% serum plus 2% BSA, which has the best blocking effect in my hand. All incubations are at room temperature. Secondary antibody incubation is always 30 min. 0.5% Triton-x100 was used for the permeablization. PBS was used as the buffer. An Ultraview confocal microscope was used for the image.
Image	Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker for ICC (Dog) Copyright image used under license

review by Andrew Smith submitted on 9 November 2004
Application:	Immunofluorescence
Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker (ab2733)
Additional Data
Notes	Immunofluorescent staining using the M6PR antibody resulted in the visualisation of large peri-nuclear structures in methanol fixed HEK293 cells. Cells were blocked and antibodies were bound in 5 % goat serum in PBS.
Dilution notes	Diluted 1:250 in 5 % goat serum / PBS

review by Marko Nykanen submitted on 24 September 2004
Application:	Immunohistochemistry (Frozen sections)
Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker (ab2733)
Additional Data
Notes	Late endosomes especially in the outermost epithelial layer were stained in the frozen semithin sections. Human mucosal epithelium was immunostained with Mannose 6 phosphate receptor ab (ab2733) as follows 1. acetone fixation -20oC 5min 2. serum block 30 min RT 3. dilution 1:10, 30 min RT 4. Dako Cytomation?s s EnVision kit for monoclonal AB was used
Dilution notes	Dilution notes: anti-mannose-6-P receptor antibody 1:10 in PBS containing 0,1% BSA and 2% horse serum

review by Darlene Shurek submitted on 11 June 2004
Application:	Immunofluorescence
Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker (ab2733)
Additional Data
Notes	Product worked extremely well, resulting in high signal and strong signal to background ratio. Very impressive.
Dilution notes	Product was used at 5ug/ml in 1% milk powder and placed over fix neuroblastoma (N2A) cell line overnight at 4C

review by Bonnie Howell submitted on 14 September 2003
Application:	Immunofluorescence
Mannose 6 Phosphate Receptor (Cation independent) antibody [2G11] - Late Endosome Marker (ab2733)
Additional Data
Notes	M6PR immunofluorescence staining resulted in larger, vesicle-like staining what would indicate localization to late endosomes/lysosomes. This antibody works great on U20S cells which have been blocked and then fixed in ice-cold methanol, but other fixations show very faint staining. Colocalization, however, may be necessary to definitively identify the area of interest due the nature of M6PR recycling.
Dilution notes	1:250 in PBS (0.5% Tween-20 and 0.5% BSA)