HRP Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [M2]

• WB

AbReview41524****

Western blot - HRP Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [M2] (AB49763)

Sample 1, 2, 3, 4, 5, 6 are the lysates from the strains expressing the tagged protein.

5% milk blocking was used for 1 hour.

All lanes:

Western blot - HRP Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [M2] (ab49763) at 1/3000 dilution

All lanes:

Saccharomyces cerevisiae Cell lysate - whole cell (S. cerevisiae cell lysate). at 10 µg

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Exposure time: 45s

This image is courtesy of an anonymous Abreview

• ELISA

AbReview13422****

ELISA - HRP Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [M2] (AB49763)

ELISA for the detection of FLAG^®-tag purified protein from Escherichia coli, using ab28947 (1/5000). Please refer to abreview for further experimental details.

Please note that in order to show a clearer image, we have brightened the original image.

This image is courtesy of an anonymous Abreview

• WB

Lab

Western blot - HRP Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [M2] (AB49763)

Western blot : Anti-ENO1 + ENO2 + ENO3 antibody [EPR12483] ab180943 staining at 1/1000 dilution, shown in green; Mouse anti-6x HisTag (ab18184) loading control staining at 1/20,000 dilution, shown in magenta.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk 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. For DDDDK-tag detection, HRP Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [M2] (ab49763) was used at 1/1000 dilution and developed using the ECL technique.

Exposure time for DDDDK tag : 0.1s.

All lanes:

Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR12483] (<a href='/en-us/products/primary-antibodies/nse-antibody-epr12483-neuronal-marker-ab180943'>ab180943</a>) at 1/1000 dilution

Lane 1:

ENO1 Recombinant Protein (His Tag) at 0.2 µg

Lane 2:

ENO2 Recombinant Protein (His Tag) at 0.2 µg

Lane 3:

ENO3 Recombinant Protein (His Tag) at 0.2 µg

Lane 4:

ENO1 Recombinant Protein (DDDDK Tag) at 0.2 µg

Lane 5:

ENO2 Recombinant Protein (DDDDK Tag) at 0.2 µg

Lane 6:

ENO3 Recombinant Protein (DDDDK Tag) at 0.2 µg

Secondary

Lanes 1 - 6:

Goat anti-Mouse 680RD at 1/20000 dilution

Lanes 1 - 6:

Goat anti-Rabbit 800CW at 1/20000 dilution

Observed band size: 50 kDa

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• WB

Lab

Western blot - HRP Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [M2] (AB49763)

Left : Anti-Inhibin beta E chain antibody ab103167 staining at 1/1000 dilution, shown in green; Mouse anti-CANX (ab238078) loading control staining at 1/20,000 dilution, shown in magenta.

Right : Rabbit Monoclonal [EPR29191-125] to Inhibin beta E chain ab322202 staining at 1/1000 dilution, shown in green; Mouse anti-CANX (ab238078) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 43 kDa in INHBE-DDDDK HEK-293T overexpression lysate with no signal observed at this size in INHBC-DDDDK HEK-293T overexpression lysate.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk 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 & Goat anti-Mouse 680RD at 1/20,000 dilution. For DDDDK-tag detection, HRP Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [M2] (ab49763) was used at 1/1000 dilution and developed using the ECL technique.

Neither ab103167 nor ab322202 cross-reacts with Inhibin beta C chain (INHBC). In Western Blot, we recommend ab322202 as the superior alternative to ab103167 for better specificity and sensitivity.

Exposure time for DDDDK tag : 2s.

Lanes 1 - 4:

Western blot - Anti-Inhibin beta E chain antibody (<a href='/en-us/products/primary-antibodies/inhibin-beta-e-chain-antibody-ab103167'>ab103167</a>) at 1/1000 dilution

Lanes 2, 4, 6 and 8:

Western blot - HRP Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [M2] (ab49763)

Lanes 5 - 8:

Western blot - Anti-Inhibin beta E chain antibody [EPR29191-125] (<a href='/en-us/products/primary-antibodies/inhibin-beta-e-chain-antibody-epr29191-125-ab322202'>ab322202</a>) at 1/1000 dilution

Lanes 1, 3, 5 and 7:

HEK-293 Mock Transfection Lysate at 20 µg

Lanes 2 and 6:

INHBE-DDDDK HEK-293T Overexpression Lysate at 20 µg

Lanes 4 and 8:

INHBC-DDDDK HEK-293T Overexpression Lysate at 20 µg

Secondary

Lanes 1 - 8:

Goat anti-Rabbit 800CW at 1/20000 dilution

Lanes 1 - 8:

Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 35 kDa

Observed band size: 43 kDa

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• WB

AbReview13187****

Western blot - HRP Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [M2] (AB49763)

Saccharomyces cerevisiae purified FLAG^®-tag protein was detected by using ab49763 (1/1000). Please refer to abreview for further experimental details.

All lanes:

Western blot - HRP Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [M2] (ab49763)

false

This image is courtesy of an anonymous Abreview