Key features and details
- Mouse monoclonal [2G11] to M6PR (cation independent)
- Suitable for: Flow Cyt, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Cow, Human
- Isotype: IgG2a
产品名称Anti-M6PR (cation independent)抗体[2G11]
参阅全部 M6PR (cation independent) 一抗
描述小鼠单克隆抗体[2G11] to M6PR (cation independent)
经测试应用适用于: Flow Cyt, ICC/IFmore details
种属反应性与反应: Mouse, Rat, Cow, Human
预测可用于: Non human primates, African green monkey不与反应: Hamster
corresponding to M6PR (cation independent).
表位This antibody is shown to recognize an epitope in the extracellular domain of Mannose 6 Phosphate Receptor.
- In Flow Cytometry, this antibody gave a positive signal in A431 cells. ICC/IF: HAP1 cells (HAP1-IGF2R knockout cells used as negative cell line)
存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
Our Abpromise guarantee covers the use of ab2733 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
|ICC/IF||Use a concentration of 1 - 10 µg/ml.|
功能Transport of phosphorylated lysosomal enzymes from the Golgi complex and the cell surface to lysosomes. Lysosomal enzymes bearing phosphomannosyl residues bind specifically to mannose-6-phosphate receptors in the Golgi apparatus and the resulting receptor-ligand complex is transported to an acidic prelyosomal compartment where the low pH mediates the dissociation of the complex. This receptor also binds IGF2. Acts as a positive regulator of T-cell coactivation, by binding DPP4.
序列相似性Belongs to the MRL1/IGF2R family.
Contains 1 fibronectin type-II domain.
结构域Contains 15 repeating units of approximately 147 AA harboring four disulfide bonds each. The most highly conserved region within the repeat consists of a stretch of 13 AA that contains cysteines at both ends.
细胞定位Lysosome membrane. Colocalized with DPP4 in internalized cytoplasmic vesicles adjacent to the cell surface.
- Information by UniProt
- 300 kDa mannose 6 phosphate receptor antibody
- 300 kDa mannose 6-phosphate receptor antibody
- Cation independent mannose 6 phosphate receptor antibody
ab2733 staining IGF2R in wild-type HAP1 cells (top panel) and IGF2R knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab2733 at 1ug/ml and ab6046 (Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
ICC/IF image of ab2733 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab2733, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
Overlay histogram showing HeLa cells stained with ab2733 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2733, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Immunofluorescent imaging of human cells (U2OS) with ab2733 confirms the specificity of this antibody, with the expected perinuclear vesicular staining of lysosomes.
IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour. Primary antibody used at 1/100 in 5% milk / 0.2% TWEEN for one hour, secondary antibody for 30 minutes. All blocking and incubation steps carried out at 37 degrees.
ab2733 positively staining formaldehyde fixed Human HEK 293 cells (red) in conjunction with goat anti mouse (Alexa 546). Nuclear staining was obtained using Hoechst.
This image is an edited version of an image received courtesy of an Abreview submitted by Kun Liu on 19 September 2005. We do not have any further information relating to this image.
ICC/IF image of ab2733 stained HepG2 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 (ab2733, 10µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) used at a 1/250 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.
ab2733 被引用在 98 文献中.
- Sapmaz A et al. USP32 regulates late endosomal transport and recycling through deubiquitylation of Rab7. Nat Commun 10:1454 (2019). PubMed: 30926795
- Koike S & Jahn R SNAREs define targeting specificity of trafficking vesicles by combinatorial interaction with tethering factors. Nat Commun 10:1608 (2019). PubMed: 30962439
- Poelaert KCK et al. Equine Herpesvirus 1 Bridles T Lymphocytes To Reach Its Target Organs. J Virol 93:N/A (2019). PubMed: 30651370
- Baranov MV et al. The Phosphoinositide Kinase PIKfyve Promotes Cathepsin-S-Mediated Major Histocompatibility Complex Class II Antigen Presentation. iScience 11:160-177 (2019). PubMed: 30612035
- Cui Y et al. Retromer has a selective function in cargo sorting via endosome transport carriers. J Cell Biol 218:615-631 (2019). PubMed: 30559172