Protein Analysis of Embryonic Lung Tissues

Embryonic lung tissues were pipetted in RIPA buffer with 1× protease inhibitor cocktail and 1× PMSF. The lysates were centrifuged at 13,200 rpm at 4°C for 15 min, then analyzed by Western blot as previously described [34 (link)]. The primary antibodies used were as follows: mouse anti-FLAG M2 (1:3,000, MilliporeSigma, Cat# F3165, RRID:AB_259529), rabbit anti-FAK (1:1,000, Cell Signaling Technology, Cat# 3285S, RRID:AB_2269034), rabbit anti-p-FAK (Tyr397) (1:1,000, Cell Signaling Technology, Cat# 3283S, RRID:AB_2173659), and mouse anti-alpha-tubulin (1:3,000, Developmental Studies Hybridoma Bank, Cat# 12G10, RRID:AB_1157911).

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Zhang K., Yao E., Chuang E., Chen B., Chuang E.Y., Volk R.F., Hofmann K.L., Zaro B, & Chuang P.T. (2022). Wnt5a–Vangl1/2 signaling regulates the position and direction of lung branching through the cytoskeleton and focal adhesions. PLoS Biology, 20(8), e3001759.

Publication 2022

mouse anti-FLAG M2 rabbit anti-FAK rabbit anti-p-FAK (Tyr397) mouse anti-alpha-tubulin

Alpha tubulin Antibodies Buffer Embryonic Hybridoma Lung Mouse Protease inhibitor Rabbit Ripa Tissues Western blot

Corresponding Organization : University of California, San Francisco

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Variable analysis

dependent variables

Protein expression levels of FLAG, FAK, and p-FAK (Tyr397)

control variables

Embryonic lung tissues
RIPA buffer with 1× protease inhibitor cocktail and 1× PMSF
Centrifugation conditions (13,200 rpm, 4°C, 15 min)
Western blot analysis as previously described [34]
Primary antibodies used: mouse anti-FLAG M2, rabbit anti-FAK, rabbit anti-p-FAK (Tyr397), and mouse anti-alpha-tubulin

controls

Positive control: Not specified
Negative control: Not specified

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