Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free (AB280571)

This data was developed using ab259362, the same antibody clone in a different buffer formulation.

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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free (AB280571)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab280571).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse hippocampus tissue staining SYNPR with ab314136 at a 1 : 8000 (0.06 ug/ml) dilution, WFS1 with ab259362 at 1 : 5000 (0.091 ug/ml) dilution and 5HT6 Receptor with ab315380 at a 1 : 1000 (0.504 ug/ml) dilution followed by secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-SYNPR (yellow; Opal™520), anti-WFS1 (green;Opal™570) and anti-5HT6 Receptor (magenta;Opal™690) on mouse hippocampus.
Panel B : anti-SYNPR staining the mossy fibers in mouse hippocampus.
Panel C : anti-WFS1 staining the pyramidal neurons of CA1 in mouse hippocampus.
Panel D : anti-5HT6 Receptor staining the pyramidal neurons in mouse hippocampus.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab314136, ab259362 and ab315380 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal^™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free (AB280571)

This data was developed using ab259362, the same antibody clone in a different buffer formulation.

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).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free (AB280571)

This data was developed using ab259362, the same antibody clone in a different buffer formulation.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free (AB280571)

This data was developed using ab259362, the same antibody clone in a different buffer formulation.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free (AB280571)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab280571).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse hippocampus tissue staining ProDynorphin with ab318966 at a 1 : 5000 (0.1 ug/ml) dilution, SYNPR with ab314136 at 1 : 8000 (0.06 ug/ml) dilution and Wolframin with ab259362 at a 1 : 5000 (0.091 ug/ml) dilution followed by secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-ProDynorphin (magenta; Opal^™570), anti-SYNPR (green; Opal^™520) and anti-Wolframin (yellow; Opal^™690) on mouse hippocampus.
Panel B : anti-ProDynorphin staining the mossy fibers of CA3 in mouse hippocampus.
Panel C : anti-SYNPR staining the mossy fibers of CA3 in mouse hippocampus.
Panel D : anti-Wolframin staining the pyramidal neurons of CA1 in mouse hippocampus.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab318966, ab314136 and ab259362 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal^™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free (AB280571)

This data was developed using ab259362, the same antibody clone in a different buffer formulation.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free (AB280571)

This data was developed using ab259362, the same antibody clone in a different buffer formulation.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free (AB280571)

This data was developed using ab259362, the same antibody clone in a different buffer formulation.

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

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free (AB280571)

This data was developed using ab259362, the same antibody clone in a different buffer formulation.

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).

• IP

Supplier Data

Immunoprecipitation - Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free (AB280571)

This data was developed using ab259362, the same antibody clone in a different buffer formulation.

WFS1 was immunoprecipitated from 0.35 mg Mouse eyeball tissue lysate 40 ug 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

All lanes:

Immunoprecipitation - Anti-WFS1 antibody [EPR23801-91] (<a href='/en-us/products/primary-antibodies/wfs1-antibody-epr23801-91-ab259362'>ab259362</a>)

Predicted band size: 100 kDa

Observed band size: 100 kDa,300 kDa

false

• WB

Lab

Western blot - Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free (AB280571)

This data was developed using ab259362, the same antibody clone in a different buffer formulation.

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

All lanes:

Western blot - Anti-WFS1 antibody [EPR23801-91] (<a href='/en-us/products/primary-antibodies/wfs1-antibody-epr23801-91-ab259362'>ab259362</a>) at 1/1000 dilution

All lanes:

Human eyeball tissue lysate at 40 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 100 kDa

Observed band size: 100 kDa,300 kDa

false

• WB

Lab

Western blot - Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free (AB280571)

This data was developed using ab259362, the same antibody clone in a different buffer formulation.

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

All lanes:

Western blot - Anti-WFS1 antibody [EPR23801-91] (<a href='/en-us/products/primary-antibodies/wfs1-antibody-epr23801-91-ab259362'>ab259362</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 100 kDa

Observed band size: 100 kDa,300 kDa

false

• WB

Lab

Western blot - Anti-WFS1 antibody [EPR23801-91] - BSA and Azide free (AB280571)

This data was developed using ab259362, the same antibody clone in a different buffer formulation.

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

All lanes:

Western blot - Anti-WFS1 antibody [EPR23801-91] (<a href='/en-us/products/primary-antibodies/wfs1-antibody-epr23801-91-ab259362'>ab259362</a>) 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:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 100 kDa

Observed band size: 100 kDa,300 kDa

false