Recombinant Anti-Tmem27 antibody [EPR17748] (ab200664)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17748] to Tmem27
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Tmem27 antibody [EPR17748]
See all Tmem27 primary antibodies -
Description
Rabbit monoclonal [EPR17748] to Tmem27 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human pancreas and fetal kidney lysates; Mouse and rat kidney lysates. IHC-P: Human and rat kidney tissues. ICC/IF: HEK293 and BxPC-3 cells. Flow Cyt (intra): BxPC-3 cells.
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General notes
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17748 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab200664 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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Flow Cyt (Intra) |
1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
1/500.
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WB |
1/1000. Detects a band of approximately 38 kDa (predicted molecular weight: 25 kDa).
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IHC-P |
1/10000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/500. |
WB
1/1000. Detects a band of approximately 38 kDa (predicted molecular weight: 25 kDa). |
IHC-P
1/10000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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Function
Regulator of SNARE complex function. Stimulator of beta cell replication. -
Tissue specificity
Kidney; collecting ducts. Pancreas; beta cells of islets. -
Sequence similarities
Belongs to the TMEM27 family. -
Post-translational
modificationsGlycosylated.
The extracellular domain is cleaved and released from the cell membrane of pancreatic beta cells. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 57393 Human
- Entrez Gene: 57394 Mouse
- Entrez Gene: 57395 Rat
- Omim: 300631 Human
- SwissProt: Q9HBJ8 Human
- SwissProt: Q9ESG4 Mouse
- SwissProt: Q9ESG3 Rat
- Unigene: 129614 Human
see all -
Alternative names
- UNQ679 antibody
- Collectrin antibody
- kidney-specific membrane protein antibody
see all
Images
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All lanes : Anti-Tmem27 antibody [EPR17748] (ab200664) at 1/1000 dilution
Lane 1 : Human fetal kidney lysate
Lane 2 : Human fetal brain lysate
Lane 3 : Human fetal heart lysate
Lane 4 : Human fetal spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 25 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsThe observed MW is consistent with what has been described in the literature PMID:16330323.
Please note lanes 2-4 are negative controls for Tmem27.
Blocking/Dilution buffer: 5% NFDM/TBST.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK293 (Human embryonic kidney) cells labeling Tmem27 with ab200664 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).
Cytoplasm staining on HEK293 cell line is observed.
The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab200664 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized BxPC-3 (Human pancreas adenocarcinoma cells) cells labeling Tmem27 with ab200664 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).
Cytoplasm staining on BxPC-3 cell line is observed.
The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab200664 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution. -
All lanes : Anti-Tmem27 antibody [EPR17748] (ab200664) at 1/1000 dilution
Lane 1 : Human pancreas lysate
Lane 2 : Human fetal brain lysate
Lane 3 : Human fetal heart lysate
Lane 4 : Human fetal spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 25 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesThe observed MW is consistent with what has been described in the literature PMID:16330323.
Please note lanes 2-4 are negative controls for Tmem27.
Blocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Tmem27 antibody [EPR17748] (ab200664) at 1/1000 dilution
Lane 1 : Mouse kidney lysate
Lane 2 : Mouse brain lysate
Lane 3 : Mouse heart lysate
Lane 4 : Mouse spleen lysate
Lane 5 : Rat kidney lysate
Lane 6 : Rat brain lysate
Lane 7 : Rat heart lysate
Lane 8 : Rat spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 25 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsThe observed MW is consistent with what has been described in the literature PMID:16330323.
Please note lanes 2-4 and 6-8 are negative controls for Tmem27.
Blocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Tmem27 with ab200664 at 1/10000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.
Luminal surface and cytoplasm staining on Human kidney tissue is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Tmem27 with ab200664 at 1/10000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.
Luminal surface and cytoplasm staining on rat kidney tissue is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling Tmem27 with ab200664 at 1/10000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.
No staining on Human cerebral cortex tissue is observed (Negative control for Tmem27).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed BxPC-3 (Human pancreas adenocarcinoma cells) cells labeling Tmem27 with ab200664 at 1/100 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730;black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab200664 has been referenced in 1 publication.
- Sarlos DP et al. Shift of Keratin Expression Profile in End-stage Kidney Increases the Risk of Tumor Development. Anticancer Res 38:5217-5222 (2018). PubMed: 30194170