HeLa nuclear extract lysate

8 Images

• WB

Unknown

Western blot - HeLa nuclear extract lysate (AB150036)

Abcam recommends using 5% milk as the blocking agent, decreasing to 2% milk during primary and secondary incubation. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

All lanes:

Western blot - Anti-HIF-1 alpha antibody [EP1215Y] (<a href='/en-us/products/primary-antibodies/hif-1-alpha-antibody-ep1215y-ab51608'>ab51608</a>) at 1/2000 dilution

Lane 1:

Western blot - HeLa nuclear extract lysate (ab150036) at 40 µg

Lane 2:

Western blot - Hela-DFO treated (0.5mM, 24h) Nuclear Lysate (<a href='/en-us/products/tissue-lysates/hela-dfo-treated-05mm-24h-nuclear-lysate-ab180880'>ab180880</a>) at 40 µg

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 1/10000 dilution

Predicted band size: 92 kDa

Observed band size: 110 kDa

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Exposure time: 8min

• WB

Unknown

Western blot - HeLa nuclear extract lysate (AB150036)

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• WB

Unknown

Western blot - HeLa nuclear extract lysate (AB150036)

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• WB

Lab

Western blot - HeLa nuclear extract lysate (AB150036)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab197874 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

All lanes:

HRP Anti-TATA binding protein TBP antibody [mAbcam 51841] - Loading Control (<a href='/en-us/products/unavailable/hrp-tata-binding-protein-tbp-antibody-mabcam-51841-loading-control-ab197874'>ab197874</a>) at 1/5000 dilution

Lane 1:

Western blot - HeLa whole cell lysate (<a href='/en-us/products/cell-lysates/hela-whole-cell-lysate-ab150035'>ab150035</a>) at 20 µg

Lane 2:

Western blot - HeLa nuclear extract lysate (ab150036) at 20 µg

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Exposure time: 1min

• WB

Unknown

Western blot - HeLa nuclear extract lysate (AB150036)

false

• WB

Unknown

Western blot - HeLa nuclear extract lysate (AB150036)

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• SDS-PAGE

Unknown

SDS-PAGE - HeLa nuclear extract lysate (AB150036)

All lanes stained with Optiblot Blue (ab119211) (20 ml)

Lane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate (ab150036)

Lysates/proteins at 10 μg per lane.

• WB

Lab

Western blot - HeLa nuclear extract lysate (AB150036)

We recommend using 5% milk in TBST as the blocking agent, decreasing to 2% milk in TBST during primary and secondary antibody incubation.

Blots were developed with Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) secondary antibody

All lanes:

Western blot - Anti-HIF-1 alpha antibody [H1alpha67] (<a href='/en-us/products/primary-antibodies/hif-1-alpha-antibody-h1alpha67-ab1'>ab1</a>) at 5 µg/mL

Lane 1:

Western blot - HeLa nuclear extract lysate (ab150036) at 40 µg

Lane 2:

Western blot - Hela-DFO treated (0.5mM, 24h) Nuclear Lysate (<a href='/en-us/products/tissue-lysates/hela-dfo-treated-05mm-24h-nuclear-lysate-ab180880'>ab180880</a>) at 40 µg

Lane 3:

HeLa nuclear control at 40 µg

Lane 4:

HeLa nuclear DFO treated at 40 µg

Lane 5:

Western blot - Recombinant Human HIF-1 alpha protein (<a href='/en-us/products/proteins-peptides/recombinant-human-hif-1-alpha-protein-ab154478'>ab154478</a>) at 0.001 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/10000 dilution

Predicted band size: 92 kDa

false

Exposure time: 20min