Anti-SEC22B antibody [EPR12335] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SEC22B antibody [EPR12335] - BSA and Azide free (AB250328)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181076). Immunocytochemistry/ Immunofluorescence analysis of SH-SY5Y (human neuroblastoma epithelial cell) cells labeling SEC22B with ab181076 at 1/50dilution (10.6 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

• IP

Supplier Data

Immunoprecipitation - Anti-SEC22B antibody [EPR12335] - BSA and Azide free (AB250328)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181076). ab181076 at 1/1000 dilution (2ug in 0.35mg lysates) immunoprecipitating SEC22B in SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate. Lane 1 (input) : SH-SY5Y whole cell lysate 10ug Lane 2 (+) : ab181076 & SH-SY5Y whole cell lysate Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab181076 in SH-SY5Y whole cell lysate For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution as a secondary antibody. Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-SEC22B antibody [EPR12335] (<a href='/en-us/products/primary-antibodies/sec22b-antibody-epr12335-ab181076'>ab181076</a>) at 1/1000 dilution

All lanes:

SH-SY5Y (human neuroblastoma epithelial cell) 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: 25 kDa

Observed band size: 28 kDa

false

Exposure time: 15s

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-SEC22B antibody [EPR12335] - BSA and Azide free (AB250328)

Intracellular Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling SEC22B with purified ab181076 at 1/50 dilution (10μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181076).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SEC22B antibody [EPR12335] - BSA and Azide free (AB250328)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling SEC22B with purified ab181076 at 1/250 dilution (2.2 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181076).

• IP

Unknown

Immunoprecipitation - Anti-SEC22B antibody [EPR12335] - BSA and Azide free (AB250328)

ab181076 (purified ) at 1/30 dilution (2ug) immunoprecipitating SEC22B in HeLa whole cell lysate.

Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+) : ab181076 & HeLa whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab181076 in HeLa whole cell lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

Blocking and diluting buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181076).

All lanes:

Immunoprecipitation - Anti-SEC22B antibody [EPR12335] (<a href='/en-us/products/primary-antibodies/sec22b-antibody-epr12335-ab181076'>ab181076</a>)

Predicted band size: 25 kDa,29 kDa

Observed band size: 32 kDa

false