Anti-SPTBN1 antibody [EPR5869] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPTBN1 antibody [EPR5869] - BSA and Azide free (AB248042)

This data was developed using ab124888, the same antibody clone in a different buffer formulation.

ab124888, at 1/250 dilution, staining SPTBN1 in paraffin-embedded Human kidney tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• WB

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Western blot - Anti-SPTBN1 antibody [EPR5869] - BSA and Azide free (AB248042)

This data was developed using ab124888, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-SPTBN1 antibody [EPR5869] (<a href='/en-us/products/primary-antibodies/sptbn1-antibody-epr5869-ab124888'>ab124888</a>) at 1/1000 dilution

Lane 1:

SH-SY5Y cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

Caco 2 cell lysate at 10 µg

Lane 4:

Jurkat cell lysate at 10 µg

Predicted band size: 275 kDa

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• WB

Lab

Western blot - Anti-SPTBN1 antibody [EPR5869] - BSA and Azide free (AB248042)

This data was developed using ab72239, the same antibody clone in a different buffer formulation.

Western blot : Anti-SPTBN1 antibody ab72239 staining at 0.04 µg/mL, shown in green; Mouse anti alpha Tubulin ab7291 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 240-260 kDa in Wild-type A549 cell lysates with no signal observed at this size in SPTBN1 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc BSA in TBS-0.1 % Tween^® 20 (TBS-T) 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 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-SPTBN1 antibody (<a href='/en-us/products/primary-antibodies/sptbn1-antibody-ab72239'>ab72239</a>) at 0.04 µg/mL

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

SPTBN1 knockout A549 at 20 µg

Lane 3:

Calu-3 at 20 µg

Lane 4:

THP-1 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 275 kDa

Observed band size: 240-260 kDa,50 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-SPTBN1 antibody [EPR5869] - BSA and Azide free (AB248042)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.