Anti-Twist2 antibody

• IP

Unknown

Immunoprecipitation - Anti-Twist2 antibody (AB66031)

Twist2 was immunoprecipitated using 0.5mg Mouse Liver tissue lysate, 5µg of Rabbit polyclonal to Twist2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Liver tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab66031.
Secondary : Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
Band : 18kDa; Twist2

All lanes:

Immunoprecipitation - Anti-Twist2 antibody (ab66031)

Predicted band size: 18 kDa

false

• WB

Unknown

Western blot - Anti-Twist2 antibody (AB66031)

This antibody is able to detect both Twist1 and Twist2 proteins. However the affinity for Twist2 is much greater. The BOI at 18kDa is related to the higher affinity binding of Twist2. In select lysates, we have observed a weaker band between 21-25kDa likely to be the cross reactivity of Twist1.

All lanes:

Western blot - Anti-Twist2 antibody (ab66031) at 1 µg/mL

Lane 1:

Human liver tissue lysate - total protein (<a href='/en-us/products/unavailable/human-liver-tissue-lysate-total-protein-ab29889'>ab29889</a>) at 10 µg

Lane 2:

Human small intestine tissue lysate - total protein (<a href='/en-us/products/unavailable/human-small-intestine-tissue-lysate-total-protein-ab29276'>ab29276</a>) at 10 µg

Lane 3:

Human testis tissue lysate - total protein (<a href='/en-us/products/unavailable/human-testis-tissue-lysate-total-protein-ab30257'>ab30257</a>) at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 18 kDa

Observed band size: 18 kDa,24 kDa,55 kDa,90 kDa

true

• WB

Lab

Western blot - Anti-Twist2 antibody (AB66031)

This antibody is able to detect both Twist1 and Twist2 proteins. However the affinity for Twist2 is much greater. The BOI at 18kDa is related to the higher affinity binding of Twist2. In select lysates, we have observed a weaker band between 21-25kDa likely to be the cross reactivity of Twist1.

All lanes:

ab66031 at 1 µg/mL

Lane 1:

Human liver tissue lysate at 10 µg

Lane 2:

Human HEK293 whole cell lysate at 10 µg

Lane 3:

Human T47D whole cell lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 3000 Cells

Predicted band size: 18 kDa

Observed band size: 21 kDa,45 kDa,46 kDa

true

Exposure time: 1min

• WB

Lab

Western blot - Anti-Twist2 antibody (AB66031)

This antibody is able to detect both Twist1 and Twist2 proteins. However the affinity for Twist2 is much greater, as demonstrated by the use of the recombinant proteins.

All lanes:

ab66031 at 1 µg/mL

Lane 1:

Recombinant Human Twist2 protein <a href='/en-us/products/unavailable/recombinant-human-twist2-protein-ab132641'>ab132641</a> at 0.1 µg

Lane 2:

Recombinant Human Twist2 protein <a href='/en-us/products/unavailable/recombinant-human-twist2-protein-ab153585'>ab153585</a> at 0.1 µg

Lane 3:

Recombinant Human Twist protein <a href='/en-us/products/unavailable/recombinant-human-twist-protein-ab112368'>ab112368</a> at 0.1 µg

Lane 4:

Recombinant Human Twist protein <a href='/en-us/products/proteins-peptides/recombinant-human-twist-protein-ab132349'>ab132349</a> at 0.1 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 18 kDa

Observed band size: 37 kDa,45 kDa,48 kDa

false

Exposure time: 4min