Anti-CAPS2 antibody [EPR27004-72] - BSA and Azide free

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CAPS2 antibody [EPR27004-72] - BSA and Azide free (AB307725)

This data was developed using ab307724, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebellum (fresh) tissue labeling CAPS2 with ab307724 at 1/100 dilution (5.2 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing positive staining on rat cerebellum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab307724 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) preadsorbed at 1/1000 dilution (2 µg/mL).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CAPS2 antibody [EPR27004-72] - BSA and Azide free (AB307725)

This data was developed using ab307724, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebellum (fresh) tissue labeling CAPS2 with ab307724 at 1/100 dilution (5.2 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing positive staining on mouse cerebellum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab307724 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) preadsorbed at 1/1000 dilution (2 µg/mL).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CAPS2 antibody [EPR27004-72] - BSA and Azide free (AB307725)

This data was developed using ab307724, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse spleen (fresh) tissue labeling CAPS2 with ab307724 at 1/100 dilution (5.2 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Negative control : confocal image showing no staining on mouse spleen (PMID : 14530279).  The nuclear counterstain was DAPI (Blue). The section was incubated with ab307724 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) preadsorbed at 1/1000 dilution (2 µg/mL).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CAPS2 antibody [EPR27004-72] - BSA and Azide free (AB307725)

This data was developed using ab307724, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat spleen (fresh) tissue labeling CAPS2 with ab307724 at 1/100 dilution (5.2 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Negative control : confocal image showing no staining on rat spleen (PMID : 14530279).  The nuclear counterstain was DAPI (Blue). The section was incubated with ab307724 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) preadsorbed at 1/1000 dilution (2 µg/mL).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CAPS2 antibody [EPR27004-72] - BSA and Azide free (AB307725)

This data was developed using ab307724, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cardiac muscle (fresh) tissue labeling CAPS2 with ab307724 at 1/100 dilution (5.2 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Negative control : confocal image showing no staining on mouse cardiac muscle (PMID : 14530279). The nuclear counterstain was DAPI (Blue). The section was incubated with ab307724 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) preadsorbed at 1/1000 dilution (2 µg/mL).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CAPS2 antibody [EPR27004-72] - BSA and Azide free (AB307725)

This data was developed using ab307724, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse skeletal muscle (fresh) tissue labeling CAPS2 with ab307724 at 1/100 dilution (5.2 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Negative control : confocal image showing no staining on mouse skeletal muscle (PMID : 14530279). The nuclear counterstain was DAPI (Blue). The section was incubated with ab307724 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) preadsorbed at 1/1000 dilution (2 µg/mL).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CAPS2 antibody [EPR27004-72] - BSA and Azide free (AB307725)

This data was developed using ab307724, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cardiac muscle (fresh) tissue labeling CAPS2 with ab307724 at 1/100 dilution (5.2 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Negative control : confocal image showing no staining on rat cardiac muscle (PMID : 14530279). The nuclear counterstain was DAPI (Blue). The section was incubated with ab307724 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) preadsorbed at 1/1000 dilution (2 µg/mL).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CAPS2 antibody [EPR27004-72] - BSA and Azide free (AB307725)

This data was developed using ab307724, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat skeletal muscle (fresh) tissue labeling CAPS2 with ab307724 at 1/100 dilution (5.2 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/mL) (Green). Negative control : confocal image showing no staining on rat skeletal muscle (PMID : 14530279). The nuclear counterstain was DAPI (Blue). The section was incubated with ab307724 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) preadsorbed at 1/1000 dilution (2 µg/mL).

• WB

Lab

Western blot - Anti-CAPS2 antibody [EPR27004-72] - BSA and Azide free (AB307725)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab307724).

Western blot : Anti-CAPS2 antibody [EPR27004-72] ab307724 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin ab7291 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 150 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in CADPS2 knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween^™ 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-CAPS2 antibody [EPR27004-72] (<a href='/en-us/products/primary-antibodies/caps2-antibody-epr27004-72-ab307724'>ab307724</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG whole cell lysate at 20 µg

Lane 2:

CADPS2 knockout U-87 MG whole cell lysate at 20 µg

Lane 3:

HUVEC whole cell lysate at 20 µg

Lane 4:

MCF7 whole cell lysate at 20 µg

Lane 5:

Raji whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 148 kDa

Observed band size: 150 kDa

false

• WB

Lab

Western blot - Anti-CAPS2 antibody [EPR27004-72] - BSA and Azide free (AB307725)

This data was developed using ab307724, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST The identity of the lower MW band at approximately 35 kDa is unknown. Exposure time : Lane 1 : 103 seconds; Lane 2 : 26 seconds.

All lanes:

Western blot - Anti-CAPS2 antibody [EPR27004-72] (<a href='/en-us/products/primary-antibodies/caps2-antibody-epr27004-72-ab307724'>ab307724</a>) at 1/1000 dilution

Lane 1:

Human brain tissue lysate at 20 µg

Lane 2:

Human cerebellum tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 150 kDa

false

• WB

Lab

Western blot - Anti-CAPS2 antibody [EPR27004-72] - BSA and Azide free (AB307725)

This data was developed using ab307724, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. This antibody does not cross-react with human CAPS1. In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution. Anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 6 seconds

All lanes:

Western blot - Anti-CAPS2 antibody [EPR27004-72] (<a href='/en-us/products/primary-antibodies/caps2-antibody-epr27004-72-ab307724'>ab307724</a>) at 1/1000 dilution

Lanes 1 - 2:

HEK-293 transfected with a human CAPS2 (WT) expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

HEK-293 transfected with a human CAPS1 (WT) expression vector containing a myc-His-tag® 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 1/100000 dilution

Observed band size: 150 kDa

false

Exposure time: 6s

• WB

Lab

Western blot - Anti-CAPS2 antibody [EPR27004-72] - BSA and Azide free (AB307725)

This data was developed using ab307724, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : spleen (PMID : 14530279, 17428348). The identity of the lower MW band at approximately 35 kDa is unknown. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : Lanes 1-4 : 37 seconds; Lanes 5-6 : 92 seconds.

All lanes:

Western blot - Anti-CAPS2 antibody [EPR27004-72] (<a href='/en-us/products/primary-antibodies/caps2-antibody-epr27004-72-ab307724'>ab307724</a>) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate 20 μg at 20 µg

Lane 2:

Rat cerebellum tissue lysate 20 μg at 20 µg

Lane 3:

Rat spleen tissue lysate at 20 µg

Lane 4:

Mouse brain tissue lysate at 20 µg

Lane 5:

Mouse cerebellum tissue lysate at 20 µg

Lane 6:

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: 150 kDa

false