Recombinant Anti-WFS1 antibody [EPR23801-91] (ab259362)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23801-91] to WFS1
- Suitable for: IP, IHC-Fr, WB, IHC-P, ICC
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-WFS1 antibody [EPR23801-91]
See all WFS1 primary antibodies -
Description
Rabbit monoclonal [EPR23801-91] to WFS1 -
Host species
Rabbit -
Tested applications
Suitable for: IP, IHC-Fr, WB, IHC-P, ICCmore details
Unsuitable for: Flow Cyt (Intra) -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse eyeball and brain tissue lysates; Rat eyeball and brain tissue lysates, Human eyeball tissue lysate; PC-12, HepG2, HeLa, HEK-293T, NIH/3T3 and PC-12 whole cell lysates. IHC-P: Mouse pancreas and cerebrum tissue; Rat pancreas and cerebrum tissue. IHC-Fr: Rat pancreas tissue; Mouse pancreas tissue. ICC: HepG2 cells. IP: Mouse eyeball tissue lysate.
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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
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23801-91 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Dyes/Markers
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab259362 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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IP |
1/30.
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IHC-Fr |
1/50.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
|
WB |
1/1000. Predicted molecular weight: 100 kDa.
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IHC-P |
1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
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ICC |
1/50.
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Notes |
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IP
1/30. |
IHC-Fr
1/50. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
WB
1/1000. Predicted molecular weight: 100 kDa. |
IHC-P
1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC
1/50. |
Target
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Relevance
WFS1 is a novel component of Wolfram syndrome, a rare form of juvenile diabetes. WFS1 plays an important role in maintaining homeostasis of the endoplasmic reticulum (ER) in the pancreas. It is normally up-regulated during insulin secretion, whereas inactivation of the protein can cause ER stress. Chronic ER stress is a major involvement in Wolfram syndrome. -
Cellular localization
Endoplasmic reticulum; endoplasmic reticulum membrane; multipass membrane protein -
Database links
- Entrez Gene: 7466 Human
- Entrez Gene: 22393 Mouse
- Entrez Gene: 83725 Rat
- Omim: 606201 Human
- SwissProt: O76024 Human
- SwissProt: P56695 Mouse
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Alternative names
- CTRCT41 antibody
- DFNA14 antibody
- DFNA38 antibody
see all
Images
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IHC image of WFS1 staining in a section of frozen normal human pancreas* performed on a Leica BOND™ system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab259362, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre -
IHC image of WFS1 staining in a section of frozen normal human hippocampus* performed on a Leica BOND™ system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab259362, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WFS1 antibody [EPR23801-91] (ab259362)
Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labelling WFS1 with ab259362 at 1/5000 dilution (0.093 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on mouse pancreatic islet (PMID: 15994758). The section was incubated with ab259362 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 cells labelling WFS1 with ab259362 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HepG2 cells is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse pancreas tissue labeling WFS1 with ab259362 at 1/50 (9.28 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse pancreatic islets is observed. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
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All lanes : Anti-WFS1 antibody [EPR23801-91] (ab259362) at 1/1000 dilution
Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : HEK-293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 4 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 5 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 100 kDa
Observed band size: 300, 100 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
The observed MW is consistent with what has been described in the literature (PMID: 12913071).
Lysates were made freshly and used in WB test immediately to minimize protein degradation.
Samples (except lane1) are non-boiled as boiling may cause protein aggregates.
Exposure time: Lane 1: 10 secondsLane 2-5: 3 minutes
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WFS1 was immunoprecipitated from 0.35 mg Mouse eyeball tissue lysate with ab259362 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259362 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Mouse eyeball tissue lysate 40 ug
Lane 2: ab259362 IP in Mouse eyeball tissue lysate
Lane 3:Rabbit monoclonal IgG (ab172730) instead of ab259362 in mouse eyeball tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 50 seconds
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Anti-WFS1 antibody [EPR23801-91] (ab259362) at 1/1000 dilution + Human eyeball tissue lysate at 40 µg
Secondary
Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 100 kDa
Observed band size: 300, 100 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
The observed MW is consistent with what has been described in the literature (PMID: 12913071 ).
Samples are non-boiled as boiling may cause protein aggregates.
Exposure time: 8 seconds
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WFS1 antibody [EPR23801-91] (ab259362)
Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling WFS1 with ab259362 at 1/5000 dilution (0.093 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on the CA1 region of mouse hippocampus (PMID: 24694561). The section was incubated with ab259362 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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All lanes : Anti-WFS1 antibody [EPR23801-91] (ab259362) at 1/1000 dilution
Lane 1 : Mouse eyeball tissue lysate at 40 µg
Lane 2 : Mouse brain tissue lysate at 40 µg
Lane 3 : Mouse spleen tissue lysate at 40 µg
Lane 4 : Rat eyeball tissue lysate at 40 µg
Lane 5 : Rat brain tissue lysate at 40 µg
Lane 6 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 100 kDa
Observed band size: 300, 100 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
The observed MW is consistent with what has been described in the literature (PMID: 12913071).
Low expression: spleen, PC-12 (PMID: 12913071; PMID: 11181571 ).
Samples are non-boiled as boiling may cause protein aggregates.
Exposure time: 8 seconds
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WFS1 antibody [EPR23801-91] (ab259362)
Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labelling WFS1 with ab259362 at 1/5000 dilution (0.093 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on rat pancreatic islet. The section was incubated with ab259362 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat pancreas tissue labeling WFS1 with ab259362 at 1/50 (4.64 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on rat pancreatic islets is observed. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WFS1 antibody [EPR23801-91] (ab259362)
Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling WFS1 with ab259362 at 1/5000 dilution (0.093 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on the CA1 region of rat hippocampus. The section was incubated with ab259362 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WFS1 antibody [EPR23801-91] (ab259362)
Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling WFS1 with ab259362 at 1/5000 dilution (0.093 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: almost no staining on mouse spleen. The section was incubated with ab259362 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab259362 has not yet been referenced specifically in any publications.