Protein samples were extracted from testes and the cultured cells using RIPA buffer and following the standard protocol. After quantification, proteins were separated by SDS-PAGE, and Western blot analysis was performed as previously described [23 (link)]. The primary antibodies used were mouse anti-UCHL1 (1: 1,000; ab8189, Abcam), rabbit anti-PLZF (1: 1,000; ab104854, Abcam), mouse anti-THY1 (1: 1,000; ab205719, Abcam), rabbit anti-VASA (1: 1,000; ab13840, Abcam), rabbit anti-DAZL (1: 500; ab34139, Abcam), goat anti-CXCR4 (1: 500; ab1670, Abcam), rabbit anti-SV40 (1: 1,000; 15,729, Cell Signaling Technology), mouse anti-PCNA (1: 1,000; sc-56, Santa Cruz Biotechnology), mouse anti-β-actin (1: 3,000; CW0096, CWBIO), rabbit anti-KIT (1: 1,000; 3074, Cell Signaling Technology) and rabbit anti-STRA8 (1: 1,000; ab49602, Abcam). The secondary antibodies used were goat anti-rabbit IgG-HRP (1: 2,000; CW0156, CWBIO), goat anti-mouse IgG-HRP (1: 3,000; CW0110, CWBIO) and rabbit anti-goat IgG-HRP (1: 3,000; CW0109, CWBIO). Finally, the blots were visualized with a Western Bright ECL Kit (Comwin) and chemiluminescence (Chemi-Doc XRS, Bio-Rad).
Cw0156
Manufactured by CWBIO
Sourced in China
The CW0156S is a laboratory equipment designed for general scientific applications. It serves as a reliable tool for researchers and scientists in various fields. The core function of the CW0156S is to provide a controlled environment for experimental procedures.
Automatically generated - may contain errors
Lab products found in correlation
Cw0110, by CWBIO (2 mentions)
Cw0102, by CWBIO (2 mentions)
Chemidoc xr, by Bio-Rad (1 mentions)
Cw0096, by CWBIO (1 mentions)
Ab1670, by Abcam (1 mentions)
Ab49602, by Abcam (1 mentions)
Ab8189, by Abcam (1 mentions)
Ab34139, by Abcam (1 mentions)
Ab104854, by Abcam (1 mentions)
Ab13840, by Abcam (1 mentions)
Ab205719, by Abcam (1 mentions)
Sc 56, by Santa Cruz Biotechnology (1 mentions)
Radioimmunoprecipitation assay buffer, by Beyotime (1 mentions)
Hoxa3, by Santa Cruz Biotechnology (1 mentions)
Trim29, by Abcam (1 mentions)
Gapdh, by Proteintech (1 mentions)
Ab194754, by Abcam (1 mentions)
Ab179693, by Abcam (1 mentions)
Ab201015, by Abcam (1 mentions)
Iseq00010, by Merck Group (1 mentions)
5 protocols using cw0156
1
Protein Expression Analysis in Testes
2
Western Blot Protein Expression Analysis
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Cell proteins were extracted with radioimmunoprecipitation assay buffer (Beyotime Institute of Biotechnology). After blocking with 8% skim milk, the membranes were incubated with the primary antibodies at 4°C overnight. After washing three times with Tris‐buffered saline with Tween 20, the membranes were incubated with the relative secondary antibody (dilution, 1:5000) for 1 hours at room temperature. The signalling of proteins was detected with an Enhanced Chemiluminescent Reagent Kit (New Cell & Molecular Biotech, Suzhou, China). The antibody information was as follows: HOXA3 (Santa Cruz, sc‐374237), ENO1 (Abcam, ab155102), SFN (Abcam, ab193667), TRIM29 (Abcam, ab108627), ARID1B (CST, 92964S), SUB1 (Abcam, ab154852), GAPDH (Proteintech, 60004‐1‐Ig), Tubulin (Proteintech, 10068‐1‐AP), goat anti‐rabbit IgG (CWBIO, CW0156) and goat anti‐mouse IgG (CWBIO, CW0110).
3
Immunohistochemical Staining of Syncytin-1, p-MEK1/2, and p-ERK1/2
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IHC staining was performed according to the standard procedures using the following antibodies: Syncytin-1 (1:200, Abcam, ab179693), p-MEK1/2 (S218 + S222) (1:200, Abcam, ab194754), and p-ERK1 (T202)/ERK2 (T185) (1:200, Abcam, ab201015) were used as primary antibodies. The secondary antibody was Biotinylated goat anti-rabbit IgG (1:1000, CWBIO, cw0156s). Staining intensity was graded as 0 (negative), 1 (weak), 2 (moderate), 3 (strong), and 4 (very strong). Positive samples were scored as 2+, 3+, or 4+. Scores of 0 and 1+ were considered negative.
4
Protein Expression Analysis in Muscle Cells
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Frozen GAS and C2C12 cells were homogenized in RIPA buffer (R0010, Solarbio) containing 1 mM PMSF (P0010, Solarbio), and protease inhibitor cocktail (539133, Merck, Rahway, NJ, USA). The protein concentration was measured using a BCA protein assay kit (P0012, Beyotime, Shanghai, China). Equivalent proteins were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinyl difluoride (PVDF) membranes (ISEQ00010, Merck). After blocking the membrane with 5% non-fat dry milk for 1.5 h, primary antibodies were proceeded overnight at 4 °C and included rabbit anti-Fgf21 (ab171941, Abcam), rabbit anti-MyHC I (22280-1-AP, Proteintech), rabbit anti-MyHC IIa (ab124937, Abcam), rabbit anti-MyHC IIb (20140-1-AP, Proteintech), and mouse anti-MyHC IIx (67299-1-Ig, Proteintech), rabbit anti-TGF-β1 (bs-0086R, Bioss, Beijing, China), rabbit anti-Smad2/3 (PA5-99539, Invitrogen), rabbit anti-p-p38 MAPK (8690, CST, MA, USA), mouse anti-Tubulin (T6199, Sigma-Aldrich, MO, USA), and mouse anti-β-actin (4967, CST). HRP-conjugated goat anti-mouse (CW0102S, CWBIO, Taizhou, China) or anti-rabbit (CW0156S, CWBIO) secondary antibodies were incubated at room temperature for 1 h. ECL (PEOO10, Solarbio) was used for enhanced chemiluminescence detection, according to the manufacturer’s instructions.
5
Protein Extraction and Western Blot Analysis
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Total protein was extracted from the iBAT and eWAT of mice using RIPA Lysis Buffer (P0013B; Beyotime) according to the manufacturer's instructions. The protein concentrations were measured with a BCA kit (P0011; Beyotime). The samples were separated by 10% SDS-PAGE and transferred to nitrocellulose membranes. The membranes were blocked with 5% non-fat dried milk in Tris-buffered saline-Tween for 1 h. Consequently, the membranes were respectively incubated with antibodies in TBST overnight at 4°C. Primary antibodies include: anti-UCP-1 (23673-1-AP; Proteintech), anti-PKA-RIIβ (sc-376778; Santa), anti-phospho-PKA-RIIβ (sc-293036; Santa), anti-FGF21 (ab171941; Abcam), anti-adiponectin (A21146; Abclonal) and anti-α-tubulin (11224-1-AP; Proteintech). Subsequently, HRP-conjugated anti-mouse IgG (CW0102S; Cwbio) or anti-rabbit IgG (CW0156S; Cwbio) was incubated for 1 h in constant temperature shaker after being washed with TBST 6 times. ECL (CW0049M; Cwbio) was used for enhanced chemiluminescence detection according to the manufacturer's instructions.
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