Anti-BAF53A antibody - ChIP Grade

• ChIP

Collaborator

ChIP - Anti-BAF53A antibody - ChIP Grade (AB3882)

Sonicated Chromatin prepared from untreated (UI) or 17beta-estradiol (E₂) treated MCF7 cells was subjected to the ChIP procedure with ab3882 to BAF53A and the immunoprecipitated chromatin was analysed in the proximal region of the estrogen-responsive pS2 promoter (as shown above) and quantified by real-time PCR (values are nomalized over inputs). The primers are designed to follow the nucleosome E (including the Estrogen Responsive Element ERE). 8 μl of ab3882 and 2x10⁶ cells were used in each ChIP experiment.

This image is courtesy of Sylvain Daujat

• WB

Project

Western blot - Anti-BAF53A antibody - ChIP Grade (AB3882)

All lanes:

Western blot - Anti-BAF53A antibody - ChIP Grade (ab3882) at 1 µg/mL

Lane 1:

Western blot - NIH/3T3 whole cell lysate (<a href='/en-us/products/cell-lysates/nih-3t3-whole-cell-lysate-ab7179'>ab7179</a>) at 10 µg

Lane 2:

MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Lane 3:

Western blot - Mouse skeletal muscle tissue lysate - total protein (<a href='/en-us/products/tissue-lysates/mouse-skeletal-muscle-tissue-lysate-total-protein-ab29711'>ab29711</a>) at 10 µg

Lane 4:

Ovary (Mouse) Tissue Lysate - normal tissue at 10 µg

Lane 5:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 47 kDa

Observed band size: 120 kDa,18 kDa,47 kDa

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• WB

Collaborator

Western blot - Anti-BAF53A antibody - ChIP Grade (AB3882)

Total extracts from two different clones of MCF7 (Human breast adenocarcinoma cell line) cells expressing a TAP tagged version of Tip60 were partially purified over IgG sepharose resin and eluted with TEV protease. Western blot was performed using a 1/1000 dilution of the BAF53A antibody.

Lane 1. TEV elution of Tip60-TAP clone #1
Lane 2. TEV elution of Tip60-TAP clone #2
Lane 3. Total extract of Tip60-TAP clone #1
Lane 4. Total extract of Tip60-TAP clone #2

NB : Tip60 is detected in the total extract due to the presence of Protein A in the TAP Tag (to which the primary and secondary abs bind). The TEV protease removes the Tag in lanes 1 and 2.

All lanes:

Western blot - Anti-BAF53A antibody - ChIP Grade (ab3882) at 1/1000 dilution

Lane 1:

TEV elution of Tip60-TAP clone 1

Lane 2:

TEV elution of Tip60-TAP clone 2

Lane 3:

Total extract of Tip60-TAP clone 1

Lane 4:

Total extract of Tip60-TAP clone 2

Predicted band size: 47 kDa

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This image is courtesy of Jacques Cote, Universite Laval, Quebec, Canada