Sciatic nerves and RSC96 cells were lysed on ice in the RIPA buffer with protease inhibitor cocktail and phosphatase inhibitor cocktail for 30 min to extract total proteins. The proteins were analyzed with a bicinchoninic acid (BCA) protein assay kit (Biosynthesis, China). 30 μg/lane (sciatic nerve) and 20 μg/lane (RSC96 cell) were used for Western blot analysis as previously described8 (link),27 (link). The primary antibodies were as follows: mouse anti-IRE1α (sc-390960; 1: 1000; Santa Cruz), rabbit anti-P-IRE1α (ab48187; 1: 2000; abcam), mouse anti-XBP1 (sc-8015; 1: 1000; Abcam), mouse anti-GRP78 (sc-376768; 1: 1000; Santa Cruz), mouse anti-GADD153 (sc-7351; 1: 500; Santa Cruz), rabbit anti-Caspase-3 (ab13847; 1: 1000; abcam), rabbit anti-Caspase-12 (om273459; 1: 1000; Omnimabs), mouse anti-Bcl-2 (sc-7382; 1: 1000; Santa Cruz), mouse anti-Bax (sc-7480; 1: 1000; Santa Cruz), mouse anti-p-JNK (sc-6254; 1: 1000; Santa Cruz), and rabbit anti-PGP9.5 (ab108986; 1: 2000; Abcam). Mouse anti-β-actin (TA-09; 1: 20000; Zhongshan Goldenbridge) served as the internal control. The secondary antibodies were goat anti-mouse IgG-HRP (ZB-2305; Zhongshan Goldenbridge) and goat anti-rabbit IgG-HRP (ZB-2301; Zhongshan Goldenbridge). Western Chemiluminescent HRP Substrate and exposed to X-film to form image. The protein bands were quantitated with Image J software27 (link).
Mouse anti gadd153
Manufactured by Santa Cruz Biotechnology
Mouse anti-GADD153 is a primary antibody that recognizes the GADD153 protein. GADD153, also known as CHOP, is a transcription factor that is induced in response to various cellular stresses. This antibody can be used to detect and study the expression of GADD153 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Ab13847, by Abcam (1 mentions)
Ab48187, by Abcam (1 mentions)
Mouse anti bcl 2, by Santa Cruz Biotechnology (1 mentions)
Ab108986, by Abcam (1 mentions)
Rabbit anti caspase 3, by Abcam (1 mentions)
Rabbit anti p ire1α, by Abcam (1 mentions)
Mouse anti p jnk, by Santa Cruz Biotechnology (1 mentions)
Mouse anti β actin, by Zhongshan Golden Bridge Biotechnology (1 mentions)
Goat anti mouse igg hrp, by Zhongshan Golden Bridge Biotechnology (1 mentions)
Goat anti rabbit igg hrp, by Zhongshan Golden Bridge Biotechnology (1 mentions)
Alexa fluor secondary antibody, by Thermo Fisher Scientific (1 mentions)
Chicken anti gfap, by Thermo Fisher Scientific (1 mentions)
2 protocols using mouse anti gadd153
1
Western Blot Analysis of ER Stress Markers
2
Immunostaining of Fixed Cells
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Fixed cells were incubated with blocking buffer (3% NDS, 0.3% Triton) for 1 h and then stained with the following primary antibodies: mouse anti-PAN (EMD Millipore Corp., MAB2300, 1/1000), chicken anti-GFAP (Thermofisher, PA1-10004, 1/2000), mouse anti-GADD153 (Santa Cruz Biotechnology, sc-7351, 1/400), goat anti-p-ERK½ (the phosphorylated-ERK, the active form of the kinase) (Santa Cruz Biotechnology, sc-16982, 1/400), rabbit anti-opn4 (ATS, AB-N39, 1/ 500), rabbit anti-opn5 (Biorbyt, orb223499, 1/500). For anti-opn4 and -opn5 stainings, cells were incubated with antibodies diluted in PBS for 2 nights at 4 °C. Otherwise, antibodies were diluted in blocking buffer; incubation was done at RT for 1 h. For revelation, cells were incubated with Alexa Fluor secondary antibodies (1:500 in PBS, Invitrogen) for 1 h at RT. For all the immunostainings, negative control experiments (without incubation with a primary antibody) were performed, in order to ensure the absence of non-specific fluorescent signal.
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