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: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-M6PR (cation independent) antibody [EPR6599]
M6PR (cation independent) 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR6599] to M6PR (cation independent) -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- 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
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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
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. -
解離定数(KD 値)
KD = 3.90 x 10 -12 M Learn more about KD -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR6599 -
アイソタイプ
IgG -
研究分野
関連製品
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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)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab124767の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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Flow Cyt (Intra) |
1/100 - 1/500.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (4) |
1/50000 - 1/200000. Detects a band of approximately 300 kDa (predicted molecular weight: 274 kDa).
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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. |
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ICC/IF | (3) |
Use a concentration of 1 µg/ml.
|
特記事項 |
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Flow Cyt (Intra)
1/100 - 1/500. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/50000 - 1/200000. Detects a band of approximately 300 kDa (predicted molecular weight: 274 kDa). |
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. |
ICC/IF
Use a concentration of 1 µg/ml. |
ターゲット情報
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機能
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. -
配列類似性
Belongs to the MRL1/IGF2R family.
Contains 1 fibronectin type-II domain. -
ドメイン
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. -
細胞内局在
Lysosome membrane. Colocalized with DPP4 in internalized cytoplasmic vesicles adjacent to the cell surface. - Information by UniProt
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参照データベース
- 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 -
別名
- 300 kDa mannose 6 phosphate receptor antibody
- 300 kDa mannose 6-phosphate receptor antibody
- Cation independent mannose 6 phosphate receptor antibody
see all
画像
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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.
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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 -
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.
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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.
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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.
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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.
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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.
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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).
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Immunocytochemistry/immunofluorescence analysis of 293T cells labelling Mannose 6 Phosphate Receptor (Cation independent) with unpurified ab124767 at dilution of 1/100.
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Intracellular 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 Fluorr® 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.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (61)
ab124767 は 61 報の論文で使用されています。
- Wang L et al. Progranulin improves neural development via the PI3K/Akt/GSK-3β pathway in the cerebellum of a VPA-induced rat model of ASD. Transl Psychiatry 12:114 (2022). PubMed: 35318322
- Yap CC et al. Dynein Is Required for Rab7-Dependent Endosome Maturation, Retrograde Dendritic Transport, and Degradation. J Neurosci 42:4415-4434 (2022). PubMed: 35474277
- Ohka S et al. The uncoating of EV71 in mature late endosomes requires CD-M6PR. Biol Open 11:N/A (2022). PubMed: 35929543
- Hertel A et al. USP32-regulated LAMTOR1 ubiquitination impacts mTORC1 activation and autophagy induction. Cell Rep 41:111653 (2022). PubMed: 36476874
- Cruz E et al. CIM6P/IGF-2 Receptor Ligands Reverse Deficits in Angelman Syndrome Model Mice. Autism Res 14:29-45 (2021). PubMed: 33108069