Anti-PMP70抗体(ab3421)
Key features and details
- Rabbit polyclonal to PMP70
- Suitable for: ICC/IF, IHC-P, Flow Cyt, WB
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
概述
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产品名称
Anti-PMP70抗体
参阅全部 PMP70 一抗 -
描述
兔多克隆抗体to PMP70 -
宿主
Rabbit -
经测试应用
适用于: ICC/IF, IHC-P, Flow Cyt, WBmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide corresponding to Rat PMP70 aa 644-659.
Sequence:NYEFKKITEDTVEFGS
(Peptide available asab4965) -
阳性对照
- WB: Rat kidney and liver tissue lysate, A431, U-2 OS, HepG2 whole cell lysates. ICC: A431 KO, A431, NIH/3T3, HMVEC, NS-1, P19, A549 whole cells. IHC-P: Human duodenum tissue.
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常规说明
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
性能
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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 or -80°C. Avoid freeze / thaw cycle. -
存储溶液
Constituents: 0.1% BSA, 99% PBS -
Concentration information loading...
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纯度
Immunogen affinity purified -
克隆
多克隆 -
同种型
IgG -
研究领域
相关产品
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Compatible Secondaries
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Conjugation kits
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Isotype control
应用
Our Abpromise guarantee covers the use of ab3421 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
应用 | Ab评论 | 说明 |
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ICC/IF | Use a concentration of 1 - 10 µg/ml. | |
IHC-P | 1/500. | |
Flow Cyt | Use at an assay dependent concentration. 0.5ug/test ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | 1/500. Detects a band of approximately 70 kDa. 70kDa band represents PMP 70 from peroxisome-enriched fractions from L cells. |
靶标
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功能
Probable transporter. The nucleotide-binding fold acts as an ATP-binding subunit with ATPase activity. -
序列相似性
Belongs to the ABC transporter superfamily. ABCD family. Peroxisomal fatty acyl CoA transporter (TC 3.A.1.203) subfamily.
Contains 1 ABC transmembrane type-1 domain.
Contains 1 ABC transporter domain. -
细胞定位
Peroxisome membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 5825 Human
- Entrez Gene: 19299 Mouse
- Entrez Gene: 25270 Rat
- Omim: 170995 Human
- SwissProt: P28288 Human
- SwissProt: P55096 Mouse
- SwissProt: P16970 Rat
- Unigene: 700576 Human
see all -
别名
- 70 kDa peroxisomal membrane protein antibody
- ABC 43 antibody
- ABC D3 antibody
see all
图片
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All lanes : Anti-PMP70 antibody (ab3421) at 1/500 dilution
Lane 1 : Rat kidney tissue lysate at 20 µg
Lane 2 : Rat liver tissue lysate at 20 µg
Lane 3 : Mouse lung tissue lysate at 20 µg
Lane 4 : A431 (Human epidermoid carcinoma cell line) whole cell lysate at 10 µg
Lane 5 : U-2 OS (Human bone osteosarcoma epithelial cell line) whole cell lysate at 10 µg -
Knockdown of PMP70 was achieved by transfecting A431 (Human epidermoid carcinoma cell line) cells with PMP70 specific siRNA. Immunofluorescence analysis was performed on A-431 cells (untransfected, panel a,d), transfected with non-specific scrambled siRNA (panels b,e) and transfected with PMP70 specific siRNA (panel c,f). Cells were fixed, permeabilized, and labelled with PMP70 Rabbit Polyclonal Antibody (5 µg/ml), followed by Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 (1:2000). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI, and Rhodamine Phalloidin (1:300) was used for cytoskeletal F-actin (red) staining. Reduction of specific signal was observed upon siRNA mediated knockdown (panel c,f) confirming specificity of the antibody to PMP70 (green). The images were captured at 60X magnification.
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Flow cytometry analysis of PMP70 in HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x106 cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab3421 at a dilution of 0.5 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
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ab3421 (2µg/ml) staining PMP70 in human duodenum using an automated system (DAKO Autostainer Plus). Using this protocol there is apical cytoplasmic staining and staining of the endoplasmic reticulum in the epithelium.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required. -
All lanes : Anti-PMP70 antibody (ab3421) at 1/1000 dilution
Lane 1 : Mouse lung tissue lysate
Lane 2 : Mouse liver tissue lysate
Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lysates/proteins at 25 µg per lane.
Observed band size: 70 kDa why is the actual band size different from the predicted? -
Immunofluorescent analysis of PMP70 (green) in NIH/3T3 (Mouse embryo fibroblast cell line) ells. The cells were permeabilized with 0.1% Triton X-100 in TBS for 15 minutes, and blocked with 3% Blocker BSA in PBS for 15 minutes at room temperature. Cells were stained with ab3421 at 10 µg/mL in blocking buffer for at least 1 hour at room temperature, and then incubated with a Goat anti-Rabbit IgG Superclonal secondary antibody, Alexa Fluor 488 conjugate at a dilution of 1:1000 for 30 minutes at room temperature (green). Nuclei (blue) were stained with Hoechst 33342 dye. Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
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Flow cytometry analysis of PMP70 in NIH/3T3 (Mouse embryo fibroblast cell line) whole cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x106 cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab3421 at a dilution of 0.5 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
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Immunofluorescence analysis of PMP70 was performed using 70% confluent log phase A431 (Human epidermoid carcinoma cell line) cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with ab3421 at 5 µg/ml in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI. F-actin (Panel c: red) was stained with Rhodamine Phalloidin. Panel d represents the merged image showing cytoplasmic (peroxisomal pattern) localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
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Flow cytometry analysis of PMP70 in HepG2 (Human liver hepatocellular carcinoma cell line) cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x106 cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab3421 at a dilution of 0.5 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
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ICC/IF image of ab3421 stained human HeLa (Human epithelial adenocarcinoma cell line) 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 (ab3421, 1 µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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). This antibody also gave a positive IF result in Hek293, HepG2 and MCF7 cells.
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Immunofluorescence analysis of PMP70 using ab3421 shows staining in HMVEC (Human microvascular endothelial cell line) cells.
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Immunofluorescence analysis of PMP70 using ab3421 shows staining in NS-1 (Mouse myeloma cell line) cells.
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Immunofluorescence analysis of PMP70 using ab3421 shows staining in P19 (Mouse embryonal carcinoma cell line) cells.
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Immunofluorescence analysis of PMP70 using ab3421 shows staining in A549 (Human lung carcinoma cell line) cells.
实验方案
文献 (32)
ab3421 被引用在 32 文献中.
- Dubreuil MM et al. Systematic Identification of Regulators of Oxidative Stress Reveals Non-canonical Roles for Peroxisomal Import and the Pentose Phosphate Pathway. Cell Rep 30:1417-1433.e7 (2020). PubMed: 32023459
- Riccio V et al. Deubiquitinating enzyme USP30 maintains basal peroxisome abundance by regulating pexophagy. J Cell Biol 218:798-807 (2019). PubMed: 30700497
- Abed M et al. The Gag protein PEG10 binds to RNA and regulates trophoblast stem cell lineage specification. PLoS One 14:e0214110 (2019). PubMed: 30951545
- Morito K et al. Gut microbial metabolites of linoleic acid are metabolized by accelerated peroxisomal ß-oxidation in mammalian cells. Biochim Biophys Acta Mol Cell Biol Lipids 1864:1619-1628 (2019). PubMed: 31351225
- Guardia CM et al. Reversible association with motor proteins (RAMP): A streptavidin-based method to manipulate organelle positioning. PLoS Biol 17:e3000279 (2019). PubMed: 31100061