Recombinant Anti-WFS1 antibody [EPR23801-91] (ab259362)

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.
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.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23801-91
Isotype

IgG
Research areas

Alternative Versions
- Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free (ab280571)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Dyes/Markers
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

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
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.

Notes
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.

Application notes

Is unsuitable for Flow Cyt (Intra).

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
Alternative names
- CTRCT41 antibody
- DFNA14 antibody
- DFNA38 antibody
- DFNA6 antibody
- DIDMOAD antibody
- WFRS antibody
- WFS antibody
- WFSL antibody
- Wolfram syndrome 1 (wolframin) antibody
- Wolfram syndrome antibody
- WOLFRAMIN antibody
- wolframin ER transmembrane glycoprotein antibody
see all

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
Immunocytochemistry - Anti-WFS1 antibody [EPR23801-91] (ab259362)

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.
Immunohistochemistry (Frozen sections) - Anti-WFS1 antibody [EPR23801-91] (ab259362)

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).
Western blot - Anti-WFS1 antibody [EPR23801-91] (ab259362)

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
Immunoprecipitation - Anti-WFS1 antibody [EPR23801-91] (ab259362)

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
Western blot - Anti-WFS1 antibody [EPR23801-91] (ab259362)

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
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
Western blot - Anti-WFS1 antibody [EPR23801-91] (ab259362)

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
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
Immunohistochemistry (Frozen sections) - Anti-WFS1 antibody [EPR23801-91] (ab259362)

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).
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
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
Anti-WFS1 antibody [EPR23801-91] (ab259362)

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.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab259362? Please let us know so that we can cite the reference in this datasheet.

ab259362 has not yet been referenced specifically in any publications.

Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Dyes/Markers

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Relevance

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (0)

Customer reviews and Q&As