Anti-SLAMF6 antibody [EPR22170]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SLAMF6 antibody [EPR22170] (AB224201)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Ramos (human Burkitt's lymphoma cell line) cells labeling SLAMF6 with ab224201 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in the Ramos cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-SLAMF6 antibody [EPR22170] (AB224201)

Flow cytometric analysis of human peripheral blood mononuclear cell (PBMC) labeling SLAMF6 with ab224201 at 1/500 dilution (right panel) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SLAMF6 antibody [EPR22170] (AB224201)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (human T cell leukemia cell line from peripheral blood) cells labeling SLAMF6 with ab224201 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in the Jurkat cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-SLAMF6 antibody [EPR22170] (AB224201)

Flow cytometric analysis of Ramos (human Burkitt's lymphoma cell line) cell line labeling SLAMF6 with ab224201 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells incubated with secondary antibody only) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

• IP

Supplier Data

Immunoprecipitation - Anti-SLAMF6 antibody [EPR22170] (AB224201)

SLAMF6 was immunoprecipitated from 0.35 mg Ramos (human Burkitt's lymphoma cell line) whole cell lysate with ab224201 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab224201 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Immunoprecipitation - Anti-SLAMF6 antibody [EPR22170] (ab224201) at 1/1000 dilution

Lane 1:

Ramos whole cell lysate 10 μg (Input) at 10 µg

Lane 2:

ab224201 at 1/30 IP in Ramos whole cell lysate (+) at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab224201 in Ramos whole cell lysate (-) at 10 µg

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

Predicted band size: 37 kDa

true

Exposure time: 10s

• IP

Supplier Data

Immunoprecipitation - Anti-SLAMF6 antibody [EPR22170] (AB224201)

SLAMF6 was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate with ab224201 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab224201 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Immunoprecipitation - Anti-SLAMF6 antibody [EPR22170] (ab224201) at 1/500 dilution

Lane 1:

Jurkat whole cell lysate (input) at 10 µg

Lane 2:

ab224201 at 1/30 IP in Jurkat whole cell lysate (+) at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab224201 in Jurkat whole cell lysate (-) at 10 µg

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 1/1000 dilution

Predicted band size: 37 kDa

true

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-SLAMF6 antibody [EPR22170] (AB224201)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SLAMF6 antibody [EPR22170] (ab224201) at 1/1000 dilution

Lane 1:

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

Lane 2:

Ramos (human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Lane 3:

Human lymph node lysate at 20 µg

Lane 4:

Human tonsil 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: 37 kDa

Observed band size: 60 kDa

false

Exposure time: 3min