Anti-PMP70 antibody (ab3421)
- Datasheet
- References (22)
- Protocols
Overview
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Product nameAnti-PMP70 antibody
See all PMP70 primary antibodies -
DescriptionRabbit polyclonal to PMP70
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Host speciesRabbit
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Tested applicationsSuitable for: ICC/IF, IHC-P, Flow Cyt, WBmore details
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Species reactivityReacts with: Mouse, Rat, Hamster, Human, Non human primates
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Immunogen
Synthetic peptide corresponding to Rat PMP70 aa 644-659.
Sequence:NYEFKKITEDTVEFGS
(Peptide available asab4965)
Properties
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FormLiquid
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Storage instructionsShipped 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.
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Storage bufferConstituents: 0.1% BSA, 99% PBS
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Concentration information loading...
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PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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Immunizing Peptide (Blocking)
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Isotype control
Applications
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.
Application | Abreviews | Notes |
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ICC/IF | 1/600. | |
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.Can be blocked with PMP70 peptide (ab4965). 70kDa band represents PMP 70 from peroxisome-enriched fractions from L cells. |
Target
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FunctionProbable transporter. The nucleotide-binding fold acts as an ATP-binding subunit with ATPase activity.
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Sequence similaritiesBelongs 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. -
Cellular localizationPeroxisome membrane.
- Information by UniProt
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Database links
- Entrez Gene: 100765314 Hamster
- Entrez Gene: 5825 Human
- Entrez Gene: 19299 Mouse
- Entrez Gene: 25270 Rat
- Omim: 170995 Human
- SwissProt: P28288 Human
- SwissProt: P55096 Mouse
- SwissProt: P16970 Rat
see all -
Alternative names
- 70 kDa peroxisomal membrane protein antibody
- ABC 43 antibody
- ABC D3 antibody
see all
Images
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All lanes : Anti-PMP70 antibody (ab3421) at 1/1000 dilution
Lane 1 : Mouse lung cell lysate
Lane 2 : Mouse liver cell lysate
Lane 3 : HepG2 cell lysate
Lysates/proteins at 25 µg per lane.
Observed band size: 70 kDa why is the actual band size different from the predicted? -
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. -
ICC/IF image of ab3421 stained human HeLa 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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Flow cytometry analysis of PMP70 in 293T 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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All lanes : Anti-PMP70 antibody (ab3421) at 1/500 dilution
Lane 1 : Rat kidney whole cell extracts at 20 µg
Lane 2 : Rat liver whole cell extracts at 20 µg
Lane 3 : Mouse lung whole cell extracts at 20 µg
Lane 4 : A431 whole cell extracts at 10 µg
Lane 5 : U2OS whole cell extracts at 10 µg -
Immunocytochemistry/Immunofluorescence analysis of PMP70 using ab3421 shows staining in HMVEC Cells.
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Flow cytometry analysis of PMP70 in HepG2 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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Immunocytochemistry/Immunofluorescence analysis of PMP70 using ab3421 shows staining in NS-1 Cells.
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Flow cytometry analysis of PMP70 in NIH-3T3 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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Immunocytochemistry/Immunofluorescence analysis of PMP70 using ab3421 shows staining in p19 Cells.
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Immunocytochemistry/Immunofluorescence analysis of PMP70 using ab3421 shows staining in A549 Cells.
Protocols
References
This product has been referenced in:
- Song J et al. Dysregulation of the NUDT7-PGAM1 axis is responsible for chondrocyte death during osteoarthritis pathogenesis. Nat Commun 9:3427 (2018). Read more (PubMed: 30143643) »
- Kazmierczak M et al. Pejvakin, a Candidate Stereociliary Rootlet Protein, Regulates Hair Cell Function in a Cell-Autonomous Manner. J Neurosci 37:3447-3464 (2017). Read more (PubMed: 28209736) »