重组Anti-M6PR (cation independent)抗体[EPR6599] (ab124767)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6599] to M6PR (cation independent)
- Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
概述
-
产品名称
Anti-M6PR (cation independent)抗体[EPR6599]
参阅全部 M6PR (cation independent) 一抗 -
描述
兔单克隆抗体[EPR6599] to M6PR (cation independent) -
宿主
Rabbit -
Tested Applications & Species
Application Species Flow Cyt HumanICC/IF HumanIHC-P MouseRatHumanWB RatHuman -
免疫原
-
阳性对照
- Jurkat, 293T, C6, PC-12, NIH/3T3 and Caco-2 cell lysates; Human papillary carcinoma, thyroid and tonsil tissue; Mouse heart, kidney, colon and spleen tissue; Rat colon tissue . ICC/IF: HAP1 WT and HAP1-IGF2R knockout cells
-
常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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.
性能
-
形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
解离常数(KD)
KD = 3.90 x 10 -12 M Learn more about KD -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
-
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR6599 -
同种型
IgG -
研究领域
相关产品
-
Alternative Versions
- Alexa Fluor® 488 Anti-M6PR (cation independent) antibody [EPR6599] - Lysosome Membrane Marker (ab184807)
- Alexa Fluor® 647 Anti-M6PR (cation independent) antibody [EPR6599] - Lysosome Membrane Marker (ab198324)
- Alexa Fluor® 405 Anti-M6PR (cation independent) antibody [EPR6599] - Lysosome Membrane Marker (ab198515)
- Alexa Fluor® 594 Anti-M6PR (cation independent) antibody [EPR6599] - Lysosome Membrane Marker (ab203437)
- Alexa Fluor® 555 Anti-M6PR (cation independent) antibody [EPR6599] - Lysosome Membrane Marker (ab203438)
- APC Anti-M6PR (cation independent) antibody [EPR6599] - Lysosome Membrane Marker (ab225131)
- PE Anti-M6PR (cation independent) antibody [EPR6599] (ab225132)
- Anti-M6PR (cation independent) antibody [EPR6599] - BSA and Azide free (ab226090)
-
Compatible Secondaries
-
Conjugation kits
-
Immunizing Peptide (Blocking)
-
Isotype control
-
Related Products
应用
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab124767 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
应用 | Species |
---|---|
Flow Cyt |
Human
|
ICC/IF |
Human
|
IHC-P |
Mouse
Rat
Human
|
WB |
Rat
Human
|
应用 | Ab评论 | 说明 |
---|---|---|
WB | (3) |
1/50000 - 1/200000. Detects a band of approximately 300 kDa (predicted molecular weight: 274 kDa).Can be blocked with M6PR (cation independent) peptide (ab169957).
|
IHC-P |
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Heat up to 98°C, below boiling, and then let cool for 10-20 min. |
|
Flow Cyt |
1/100 - 1/500.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
ICC/IF | (3) |
Use a concentration of 1 µg/ml.
|
说明 |
---|
WB
1/50000 - 1/200000. Detects a band of approximately 300 kDa (predicted molecular weight: 274 kDa).Can be blocked with M6PR (cation independent) peptide (ab169957). |
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Heat up to 98°C, below boiling, and then let cool for 10-20 min. |
Flow Cyt
1/100 - 1/500. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use a concentration of 1 µg/ml. |
靶标
-
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
-
数据库链接
- Entrez Gene: 3482 Human
- Entrez Gene: 16004 Mouse
- Entrez Gene: 25151 Rat
- Omim: 147280 Human
- SwissProt: P11717 Human
- SwissProt: Q07113 Mouse
- Unigene: 487062 Human
- Unigene: 673278 Human
see all -
别名
- 300 kDa mannose 6 phosphate receptor antibody
- 300 kDa mannose 6-phosphate receptor antibody
- Cation independent mannose 6 phosphate receptor antibody
see all
图片
-
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: M6PR (cation independent) knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: A549 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab124767 observed at 274 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab124767 was shown to specifically react with M6PR (cation independent) in wild-type HAP1 cells as signal was lost in M6PR (cation independent) knockout cells. Wild-type and M6PR (cation independent) knockout samples were subjected to SDS-PAGE. Ab124767 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/50000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
-
Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)
ab124767 staining M6PR in wild-type HAP1 cells (top panel) and IGF2R knockout HAP1 cells (bottom panel). The cells were fixed with 100% MeOH for 5 min. , 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 ab124767 at 1μg/ml and ab195889 (Mouse monoclonal [DM1A] to alpha Tubulin - Microtubule Marker (Alexa Fluor® 594)) at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). -
All lanes : Anti-M6PR (cation independent) antibody [EPR6599] (ab124767) at 1/50000 dilution (purified)
Lane 1 : C6 (rat glioma) whole cell lysate
Lane 2 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysates
Lane 3 : NIH/3T3 (mouse embryo) whole cell lysate
Lane 4 : Mouse heart tissue lysate
Lane 5 : Mouse kidney tissue lysate
Lane 6 : Mouse spleen tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 274 kDa
Observed band size: 300 kDa why is the actual band size different from the predicted?
Blocking and diluting buffer 5% NFDM/TBST -
All lanes : Anti-M6PR (cation independent) antibody [EPR6599] (ab124767) at 1/50000 dilution (unpurified)
Lane 1 : Jurkat cell lysate
Lane 2 : 293T cell lysate
Lane 3 : Caco-2 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-Rabbit HRP at 1/2000 dilution
Predicted band size: 274 kDa -
All lanes : Anti-M6PR (cation independent) antibody [EPR6599] (ab124767) at 1/200000 dilution (purified)
Lane 1 : Jurkat (human acute T cell leukemia) whole cell lysate
Lane 2 : HEK293 (human embryonic kidney) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 274 kDa
Observed band size: 300 kDa why is the actual band size different from the predicted?
Blocking and diluting buffer 5% NFDM/TBST -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)
Immunohistochemical staining of paraffin embedded rat colon tissue section labelling M6PR with purified ab124767 at dilution of 1/500. The secondary antibody used was ab97051 Goat Anti-Rabbit IgG H&L (HRP), at a dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)
Immunohistochemical staining of paraffin embedded mouse colon tissue section labelling M6PR with purified ab124767 at dilution of 1/500. The secondary antibody used was ab97051 Goat Anti-Rabbit IgG H&L (HRP), at a dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)
Immunohistochemical staining of paraffin embedded human thyroid carcinoma tissue section labelling M6PR with purified ab124767 at dilution of 1/500. The secondary antibody used was ab97051 Goat Anti-Rabbit IgG H&L (HRP), at a dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)
Immunohistochemical analysis of formalin fixed, paraffin embedded Human papillary carcinoma tissue section labelling Mannose 6 Phosphate Receptor (Cation independent) with unpurified ab124767 at dilution of 1/250.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)
Immunohistochemical analysis of formalin fixed, paraffin embedded Human tonsil tissue section labelling Mannose 6 Phosphate Receptor (Cation independent) with unpurified ab124767 at dilution of 1/250.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)
Immunocytochemistry/Immunofluorescence analysis of Jurkat (human acute T cell leukemia) cells labelling M6PR with purified ab124767 at 1/100. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/1000) using ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary. Nuclei couterstained with DAPI (blue).
For negative control 1, rabbit primary antibody was used, followed by anti-mouse secondary antibody (ab150120). For negative control 2, mouse primary antibody (ab7291) was used followed by anti-rabbit secondary antibody (ab150077).
-
Immunocytochemistry/ Immunofluorescence - Anti-M6PR (cation independent) antibody [EPR6599] (ab124767)
Immunocytochemistry/immunofluorescence analysis of 293T cells labelling Mannose 6 Phosphate Receptor (Cation independent) with unpurified ab124767 at dilution of 1/100.
-
Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labelling M6PR with purified ab124767 at 1/150 (red). Cells were fixed with 4% paraformaldehyde. Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
实验方案
数据表及文件
-
SDS download
-
Datasheet download
文献 (39)
ab124767 被引用在 39 文献中.
- Curtis ME et al. Dysregulation of the Retromer Complex System in Down Syndrome. Ann Neurol 88:137-147 (2020). PubMed: 32320094
- Han J et al. Involvement of CASP9 (caspase 9) in IGF2R/CI-MPR endosomal transport. Autophagy N/A:1-17 (2020). PubMed: 32397873
- Cui L et al. The Lysosomal Membrane Protein Lamp2 Alleviates Lysosomal Cell Death by Promoting Autophagic Flux in Ischemic Cardiomyocytes. Front Cell Dev Biol 8:31 (2020). PubMed: 32117965
- Boivin M et al. Reduced autophagy upon C9ORF72 loss synergizes with dipeptide repeat protein toxicity in G4C2 repeat expansion disorders. EMBO J 39:e100574 (2020). PubMed: 31930538
- Li JG et al. A pharmacological chaperone improves memory by reducing Aß and tau neuropathology in a mouse model with plaques and tangles. Mol Neurodegener 15:1 (2020). PubMed: 31964406