Anti-SLC5A7 antibody [EPR25152-16] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-SLC5A7 antibody [EPR25152-16] - BSA and Azide free (AB289637)

This data was developed using ab289635, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized SH-SY5Y (Human neuroblastoma epithelial cell line) cells labeling SLC5A7 with ab289635 at 1/500 dilution (Red) compared with Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) was used as secondary antibody at 1/2000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SLC5A7 antibody [EPR25152-16] - BSA and Azide free (AB289637)

This data was developed using ab289635, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y cells labeling SLC5A7 with ab289635 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in SH-SY5Y cell line. The nuclear counterstain is DAPI (Blue). Tubulin is labeled using ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (Red).

Secondary antibody only control : Used PBS instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SLC5A7 antibody [EPR25152-16] - BSA and Azide free (AB289637)

This data was developed using ab289635, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C8-D1A cells labeling SLC5A7 with ab289635 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in C8-D1A cell line. The nuclear counterstain is DAPI (Blue). Tubulin is labeled using ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (Red).

Secondary antibody only control : Used PBS instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody at 1/1000 dilution.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-SLC5A7 antibody [EPR25152-16] - BSA and Azide free (AB289637)

This data was developed using ab289635, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized C8-D1A (Mouse brain astrocyte cell line) cells labeling SLC5A7 with ab289635 at 1/500 dilution (Red) compared with Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) was used as secondary antibody at 1/2000 dilution.

• WB

Lab

Western blot - Anti-SLC5A7 antibody [EPR25152-16] - BSA and Azide free (AB289637)

This data was developed using ab289635, the same antibody clone in a different buffer formulation.

5% NFDM/TBST was used as a blocking and concentration buffer.

Samples are non-boiled as boiling may cause protein aggregates.
The molecular weight observed is consistent with what has been described in the literature (PMID : 10649566).

All lanes:

Western blot - Anti-SLC5A7 antibody [EPR25152-16] (<a href='/en-us/products/primary-antibodies/slc5a7-antibody-epr25152-16-ab289635'>ab289635</a>) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human spinal cord tissue lysate at 20 µg

Lane 3:

Human liver tissue lysate at 40 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/5000 dilution

Predicted band size: 63 kDa

Observed band size: 120 kDa,63 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-SLC5A7 antibody [EPR25152-16] - BSA and Azide free (AB289637)

This data was developed using ab289635, the same antibody clone in a different buffer formulation.

Samples are non-boiled as boiling may cause protein aggregates.

5% NFDM/TBST was used as a blocking and dilution buffer.

All lanes:

Western blot - Anti-SLC5A7 antibody [EPR25152-16] (<a href='/en-us/products/primary-antibodies/slc5a7-antibody-epr25152-16-ab289635'>ab289635</a>) at 1/1000 dilution

Lane 1:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

A-172 (human brain glioblastoma) 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 1/200000 dilution

Predicted band size: 63 kDa

Observed band size: 63 kDa

false

Exposure time: 10s

• WB

Unknown

Western blot - Anti-SLC5A7 antibody [EPR25152-16] - BSA and Azide free (AB289637)

This data was developed using ab289635, the same antibody clone in a different buffer formulation.

Lysate was made freshly and used in WB test immediately to minimize protein degradation.
The molecular weight observed is consistent with what has been described in the literature (PMID : 10649566).

5% NFDM/TBST was used as a blocking and dilution buffer.

All lanes:

Western blot - Anti-SLC5A7 antibody [EPR25152-16] (<a href='/en-us/products/primary-antibodies/slc5a7-antibody-epr25152-16-ab289635'>ab289635</a>) at 1/1000 dilution

Lane 1:

C6 (Rat glial tumor glial 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 1/100000 dilution

Predicted band size: 63 kDa

Observed band size: 120 kDa,63 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-SLC5A7 antibody [EPR25152-16] - BSA and Azide free (AB289637)

This data was developed using ab289635, the same antibody clone in a different buffer formulation.

5% NFDM/TBST was used as a blocking and dilution buffer.

Lysate was made freshly and used in WB test immediately to minimize protein degradation.

All lanes:

Western blot - Anti-SLC5A7 antibody [EPR25152-16] (<a href='/en-us/products/primary-antibodies/slc5a7-antibody-epr25152-16-ab289635'>ab289635</a>) at 1/1000 dilution

All lanes:

C8-D1A (Mouse brain astrocyte) 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: 63 kDa

Observed band size: 63 kDa

false

Exposure time: 59s