Anti-SLP-2 antibody [EPR18718] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-SLP-2 antibody [EPR18718] - BSA and Azide free (AB251097)

This data was developed using ab191883, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling SLP-2 with ab191883 at 1/120 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A]-Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor^® 488) at 1/500 dilution was used as the secondary antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SLP-2 antibody [EPR18718] - BSA and Azide free (AB251097)

This data was developed using ab191883, the same antibody clone in a different buffer formulation.

Immunofluorescence staining of HeLa cells with ab191883 at a working dilution of 1 in 250, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1 in 1000 (shown in green). The mitochondria were stained with ab33985 (mouse monoclonal to COX-IV, mitochondrial marker) at 1/1000 and ab150120 at 1/1000 (shown in red). The cells were fixed in 100% methanol and permeabilized using 0.1% triton-X 100. The negative controls are shown in bottom panels. For negative control 1, ab191883 was used at a dilution of 1/250 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/1000. For negative control 2, ab7291 (mouse monoclonal to alpha tubulin) was used at a dilution of 1/1000 followed by ab150077 (Alexa Fluor^® 488 goat anti-rabbit) at a dilution of 1/1000.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SLP-2 antibody [EPR18718] - BSA and Azide free (AB251097)

This data was developed using ab191883, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized Jurkat cells (Human T cell leukemia cell line from peripheral blood) labeling SLP-2 with ab191883 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Jurkat cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin -Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows : -
-ve control 1 : ab191883 at 1/250 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SLP-2 antibody [EPR18718] - BSA and Azide free (AB251097)

This data was developed using ab191883, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling SLP-2 with ab191883 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin -Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows : -
-ve control 1 : ab191883 at 1/250 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• IP

Unknown

Immunoprecipitation - Anti-SLP-2 antibody [EPR18718] - BSA and Azide free (AB251097)

This data was developed using ab191883, the same antibody clone in a different buffer formulation.

SLP-2 was immunoprecipitated from 1mg of Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate with ab191883 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab191883 at 1/1000 dilution. Veriblot for IP (HRP) (ab13136) was used as secondary antibody at 1/10000 dilution.
Lane 1 : Jurkat whole cell lysate 10μg (Input).
Lane 2 : ab191883 IP in Jurkat whole cell lysate.
Lane 3 : Rabbit IgG,monoclonal [EPR25A] (ab172730) instead of ab191883 in Jurkat whole cell cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-SLP-2 antibody [EPR18718] (<a href='/en-us/products/primary-antibodies/slp-2-antibody-epr18718-ab191883'>ab191883</a>)

Predicted band size: 39 kDa

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• WB

Lab

Western blot - Anti-SLP-2 antibody [EPR18718] - BSA and Azide free (AB251097)

This data was developed using ab191883, the same antibody clone in a different buffer formulation.

Western blot : Anti-SLP-2 antibody [EPR18718] ab191883 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 40 kDa in Wild-type A549 cell lysates with no signal observed at this size in STOML2 knockout A549 cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 & Goat anti-Mouse 680RD at 1/20,000 dilution.

Lanes 1 - 4:

Western blot - Anti-SLP-2 antibody [EPR18718] (<a href='/en-us/products/primary-antibodies/slp-2-antibody-epr18718-ab191883'>ab191883</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-SLP-2 antibody [EPR18718] - BSA and Azide free (ab251097) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

STOML2 knockout A549 at 20 µg

Lane 3:

HeLa at 20 µg

Lane 4:

PC-3 at 20 µg

Secondary

All lanes:

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

Predicted band size: 40 kDa

Observed band size: 40 kDa

false

• WB

Supplier Data

Western blot - Anti-SLP-2 antibody [EPR18718] - BSA and Azide free (AB251097)

This data was developed using ab191883, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SLP-2 antibody [EPR18718] (<a href='/en-us/products/primary-antibodies/slp-2-antibody-epr18718-ab191883'>ab191883</a>) at 1/1000 dilution

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

A549 (Human lung carcinoma cell line) 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

Predicted band size: 39 kDa

Observed band size: 39 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-SLP-2 antibody [EPR18718] - BSA and Azide free (AB251097)

This data was developed using ab191883, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SLP-2 antibody [EPR18718] (<a href='/en-us/products/primary-antibodies/slp-2-antibody-epr18718-ab191883'>ab191883</a>) at 1/1000 dilution

All lanes:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 39 kDa

false

Exposure time: 1min

• WB

Supplier Data

Western blot - Anti-SLP-2 antibody [EPR18718] - BSA and Azide free (AB251097)

This data was developed using ab191883, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times : Lanes 1-4 : 3 minutes; Lanes 5-6 : 1 minute.

All lanes:

Western blot - Anti-SLP-2 antibody [EPR18718] (<a href='/en-us/products/primary-antibodies/slp-2-antibody-epr18718-ab191883'>ab191883</a>) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse spleen lysate at 10 µg

Lane 3:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Lane 4:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 5:

Rat brain lysate at 10 µg

Lane 6:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 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

Predicted band size: 227 kDa,39 kDa

Observed band size: 39 kDa

false