Anti-SLC26A2/DTD antibody [EPR27119-17] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-SLC26A2/DTD antibody [EPR27119-17] - BSA and Azide free (AB308626)

This data was developed using ab308625, the same antibody clone in a different buffer formulation.

SLC26A2/DTD was immunoprecipitated from 0.35 mg Human colon tissue lysate with ab308625 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab308625 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Human colon tissue lysate
Lane 2 : ab308625 IP in Human colon tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab308625 in human colon tissue lysate

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

Exposure time : 180 seconds

Bands around 300 kDa may be SLC26A2 oligomers.

All lanes:

Immunoprecipitation - Anti-SLC26A2/DTD antibody [EPR27119-17] (<a href='/en-us/products/primary-antibodies/slc26a2-dtd-antibody-epr27119-17-ab308625'>ab308625</a>) at 1/1000 dilution

All lanes:

Human colon tissue 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

Observed band size: 100 kDa,300 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-SLC26A2/DTD antibody [EPR27119-17] - BSA and Azide free (AB308626)

This data was developed using 308625, the same antibody clone in a different buffer formulation.  Blocking and diluting buffer and concentration : 5% NFDM/TBST. SLC26A2 is a glycoprotein of approximately 100 kDa and detected as a 80 kDa band after treated with Protein Deglycosylation MIX II. Samples are non-boiled as boiling may cause protein aggregation. Exposure time : 37 seconds

All lanes:

Western blot - Anti-SLC26A2/DTD antibody [EPR27119-17] (<a href='/en-us/products/primary-antibodies/slc26a2-dtd-antibody-epr27119-17-ab308625'>ab308625</a>) at 1/1000 dilution

Lane 1:

Untreated human colon tissue lysate at 20 µg

Lane 2:

Human colon tissue lysate treated with Protein Deglycosylation MIXII at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 81 kDa

Observed band size: 100 kDa,80 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-SLC26A2/DTD antibody [EPR27119-17] - BSA and Azide free (AB308626)

This data was developed using 308625, the same antibody clone in a different buffer formulation.  Blocking and diluting buffer and concentration : 5% NFDM/TBST. Low expression : Liver tissue (PMID : 24302720, PMID : 11457925) Stomach tissue. Samples are non-boiled as boiling may cause protein aggregation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 26 seconds

All lanes:

Western blot - Anti-SLC26A2/DTD antibody [EPR27119-17] (<a href='/en-us/products/primary-antibodies/slc26a2-dtd-antibody-epr27119-17-ab308625'>ab308625</a>) at 1/1000 dilution

Lane 1:

Human colon tissue lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Lane 3:

Human stomach tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 81 kDa

Observed band size: 100 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-SLC26A2/DTD antibody [EPR27119-17] - BSA and Azide free (AB308626)

This data was developed using 308625, the same antibody clone in a different buffer formulation.  Blocking and diluting buffer and concentration : 5% NFDM/TBST. Low expression : 293T. Samples are non-boiled as boiling may cause protein aggregation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 10 seconds

All lanes:

Western blot - Anti-SLC26A2/DTD antibody [EPR27119-17] (<a href='/en-us/products/primary-antibodies/slc26a2-dtd-antibody-epr27119-17-ab308625'>ab308625</a>) at 1/1000 dilution

Lane 1:

A375 (human malignant melanoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 81 kDa

Observed band size: 80 kDa,100 kDa

false

Exposure time: 10s

• WB

Lab

Western blot - Anti-SLC26A2/DTD antibody [EPR27119-17] - BSA and Azide free (AB308626)

This data was developed using ab308625, the same antibody clone in a different buffer formulation.

Western blot : Anti-SLC26A2 antibody [EPR27119-17] (ab308625) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab308625 was shown to bind specifically to SLC26A2. A band was observed at 82 kDa in wild-type A549 cell lysates with no signal observed at this size in SLC26A2 knockout cell line. To generate this image, wild-type and SLC26A2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution. For optimal results, we recommend using fluorescent western blot (TBS-based) blocking solution and not boiling the samples.

All lanes:

Western blot - Anti-SLC26A2/DTD antibody [EPR27119-17] (<a href='/en-us/products/primary-antibodies/slc26a2-dtd-antibody-epr27119-17-ab308625'>ab308625</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

SLC26A2 knockout A549 cell lysate at 20 µg

Lane 3:

HeLa UNBOILED cell lysate at 20 µg

Lane 4:

HEK-293 UNBOILED cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 82 kDa

false