Anti-FYCO1 antibody [EPR28361-79] - BSA and Azide free

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-FYCO1 antibody [EPR28361-79] - BSA and Azide free (AB316013)

This data was developed using ab316012, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum (fresh frozen) tissue labeling FYCO1 with ab316012 at 1/100 (5.16 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

Low expression : confocal image showing no staining on mouse cerebrum (PMID : 34215815). The nuclear counterstain was DAPI (Blue). The section was incubated with ab316012 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 (2 ug/mL) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FYCO1 antibody [EPR28361-79] - BSA and Azide free (AB316013)

This data was developed using ab316012, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling FYCO1 with ab316012 at 1/2000 (0.258 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat cardiac muscle. The section was incubated with ab316012 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed 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-FYCO1 antibody [EPR28361-79] - BSA and Azide free (AB316013)

This data was developed using ab316012, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling FYCO1 with ab316012 at 1/2000 (0.258 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cardiac muscle (PMID : 34215815; PMID : 33997522). The section was incubated with ab316012 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed 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-FYCO1 antibody [EPR28361-79] - BSA and Azide free (AB316013)

This data was developed using ab316012, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling FYCO1 with ab316012 at 1/2000 (0.258 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse testis. The section was incubated with ab316012 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed 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-FYCO1 antibody [EPR28361-79] - BSA and Azide free (AB316013)

This data was developed using ab316012, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling FYCO1 with ab316012 at 1/2000 (0.258 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat testis. The section was incubated with ab316012 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-FYCO1 antibody [EPR28361-79] - BSA and Azide free (AB316013)

This data was developed using ab316012, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse heart (fresh frozen) tissue labeling FYCO1 with ab316012 at 1/100 (5.16 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green).

Confocal image showing positive staining on mouse heart. The nuclear counterstain was DAPI (Blue). The section was incubated with ab316012 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 (2 ug/mL) dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-FYCO1 antibody [EPR28361-79] - BSA and Azide free (AB316013)

This data was developed using ab316012, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

FYCO1 was immunoprecipitated from 0.35 mg Mouse skeletal muscle tissue lysate with ab316012 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316012 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse skeletal muscle tissue lysate
Lane 2 : ab316012 IP in Mouse skeletal muscle tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab316012 in mouse skeletal muscle tissue lysate.

All lanes:

Immunoprecipitation - Anti-FYCO1 antibody [EPR28361-79] (<a href='/en-us/products/primary-antibodies/fyco1-antibody-epr28361-79-ab316012'>ab316012</a>) at 1/30 dilution

All lanes:

Mouse skeletal muscle tissue lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-FYCO1 antibody [EPR28361-79] - BSA and Azide free (AB316013)

This data was developed using ab316012, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : brain (PMID : 34215815).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-FYCO1 antibody [EPR28361-79] (<a href='/en-us/products/primary-antibodies/fyco1-antibody-epr28361-79-ab316012'>ab316012</a>) at 1/1000 dilution

Lane 1:

Rat heart tissue lysate at 20 µg

Lane 2:

Rat skeletal muscle tissue lysate at 20 µg

Lane 3:

Rat brain tissue 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 1/20000 dilution

Observed band size: 162 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-FYCO1 antibody [EPR28361-79] - BSA and Azide free (AB316013)

This data was developed using ab316012, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : Neuro-2a.

The lysate of lane 1 was stored at -80℃ prior to Western Blotting. The band beneath the target band (162 kDa) are likey to be degradation products.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-FYCO1 antibody [EPR28361-79] (<a href='/en-us/products/primary-antibodies/fyco1-antibody-epr28361-79-ab316012'>ab316012</a>) at 1/1000 dilution

Lane 1:

Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Neuro-2a (mouse neuroblastoma neuroblast) 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 1/20000 dilution

Observed band size: 162 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-FYCO1 antibody [EPR28361-79] - BSA and Azide free (AB316013)

This data was developed using ab316012, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : brain (PMID : 34215815), spleen (PMID : 34215815).

In lane 1-4, the lysates were stored at -80℃ prior to Western Blotting. The bands beneath the target band (162 kDa) are likey to be degradation products. In lanes 5-7, the lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-4 : 180 seconds, lanes 5-7 : 48 seconds.

All lanes:

Western blot - Anti-FYCO1 antibody [EPR28361-79] (<a href='/en-us/products/primary-antibodies/fyco1-antibody-epr28361-79-ab316012'>ab316012</a>) at 1/1000 dilution

Lane 1:

Mouse skeletal muscle tissue lysate at 20 µg

Lanes 2 and 5:

Mouse heart tissue lysate at 20 µg

Lanes 3 and 6:

Mouse brain tissue lysate at 20 µg

Lanes 4 and 7:

Mouse spleen tissue 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 1/20000 dilution

Observed band size: 162 kDa,36 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FYCO1 antibody [EPR28361-79] - BSA and Azide free (AB316013)

This data was developed using ab316012, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling FYCO1 with ab316012 at 1/2000 (0.258 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : nearly negative staining on rat cerebrum. The section was incubated with ab316012 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed 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-FYCO1 antibody [EPR28361-79] - BSA and Azide free (AB316013)

This data was developed using ab316012, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling FYCO1 with ab316012 at 1/2000 (0.258 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : nearly negative staining on mouse cerebrum (PMID : 34215815). The section was incubated with ab316012 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins