Anti-SPTLC1 antibody [EPR26226-136] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPTLC1 antibody [EPR26226-136] - BSA and Azide free (AB307553)

This data was developed using ab307552, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human colon tissue labeling SPTLC1 with ab307552 at 1/200 dilution (2.44 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human colon (PMID : 12704216). The section was incubated with ab307552 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SPTLC1 antibody [EPR26226-136] - BSA and Azide free (AB307553)

This data was developed using ab307552, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde-fixed 90% methanol-permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labeling SPTLC1 with ab307552 at 1/50 dilution (1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPTLC1 antibody [EPR26226-136] - BSA and Azide free (AB307553)

This data was developed using ab307552, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling SPTLC1 with ab307552 at 1/200 dilution (2.44 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human cerebrum. The section was incubated with ab307552 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPTLC1 antibody [EPR26226-136] - BSA and Azide free (AB307553)

This data was developed using ab307552, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling SPTLC1 with ab307552 at 1/200 dilution (2.44 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human placenta. The section was incubated with ab307552 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

• IP

Supplier Data

Immunoprecipitation - Anti-SPTLC1 antibody [EPR26226-136] - BSA and Azide free (AB307553)

This data was developed using ab307552, the same antibody clone in a different buffer formulation.

SPTLC1 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate 10 µg with ab307552 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307552 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa whole cell lysate

Lane 2 : ab307552 at 1/30 IP in HeLa whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307552 in HeLa whole cell lysate

Blocking and dilution 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-SPTLC1 antibody [EPR26226-136] (<a href='/en-us/products/primary-antibodies/sptlc1-antibody-epr26226-136-ab307552'>ab307552</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) at 10 µg

Lane 2:

<a href='/en-us/products/primary-antibodies/sptlc1-antibody-epr26226-136-ab307552'>ab307552</a> at 1/30 IP in HeLa whole cell lysate

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

Observed band size: 53 kDa

true

Exposure time: 180s

• WB

Lab

Western blot - Anti-SPTLC1 antibody [EPR26226-136] - BSA and Azide free (AB307553)

This data was developed using ab307552, the same antibody clone in a different buffer formulation.

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

Compared with ab307552, ab315158 has higher sensitivity. We recommend ab315158 as an alternative.

Lanes 1 - 4:

Western blot - Anti-SPTLC1 antibody [EPR26226-136] (<a href='/en-us/products/primary-antibodies/sptlc1-antibody-epr26226-136-ab307552'>ab307552</a>) at 1/1000 dilution

Lanes 5 - 8:

Western blot - Anti-SPTLC1 antibody [EPR26226-221] (<a href='/en-us/products/primary-antibodies/sptlc1-antibody-epr26226-221-ab315158'>ab315158</a>) at 1/1000 dilution

Lanes 1 and 5:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lanes 2 and 6:

293T (human embryonic kidney epithelial cell) whole cell lysate

Lanes 3 and 7:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lanes 4 and 8:

Human small intestine 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: 54 kDa

false

Exposure time: 60s

• WB

Supplier Data

Western blot - Anti-SPTLC1 antibody [EPR26226-136] - BSA and Azide free (AB307553)

This data was developed using ab307552, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : human skeletal muscle (PMID : 17023427). Exposure time : 48 seconds.

All lanes:

Western blot - Anti-SPTLC1 antibody [EPR26226-136] (<a href='/en-us/products/primary-antibodies/sptlc1-antibody-epr26226-136-ab307552'>ab307552</a>) at 1/1000 dilution

Lane 1:

Human placenta tissue lysate at 20 µg

Lane 2:

Human skeletal muscle 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: 53 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-SPTLC1 antibody [EPR26226-136] - BSA and Azide free (AB307553)

This data was developed using ab307552, 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 SPTLC2 and SPTLC3. Exposure time : 37 seconds.

All lanes:

Western blot - Anti-SPTLC1 antibody [EPR26226-136] (<a href='/en-us/products/primary-antibodies/sptlc1-antibody-epr26226-136-ab307552'>ab307552</a>) at 1/1000 dilution

Lane 1:

His-tagged human SPTLC1 recombinant protein fragment at 10 ng

Lane 2:

His-tagged human SPTLC2 recombinant protein fragment at 10 ng

Lane 3:

His-tagged human SPTLC3 recombinant protein fragment at 10 ng

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: 46 kDa

false

Exposure time: 37s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPTLC1 antibody [EPR26226-136] - BSA and Azide free (AB307553)

This data was developed using ab307552, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling SPTLC1 with ab307552 at 1/200 dilution (2.44 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Negative control : No staining on human skeletal muscle. The section was incubated with ab307552 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).