Ceramide synthase 2, also known as LAG1 longevity assurance homolog 2 or Tumor metastasis-suppressor gene 1 protein is an enzyme that in humans is encoded by the CERS2 gene. Ceramide synthase 2 is a ceramide synthase that catalyses the synthesis of very long acyl chain ceramides, including C20 and C26 ceramides. It is the most ubiquitously expressed of all CerS and has the broadest distribution in the human body. CerS2 was first identified in 2001. It contains the conserved TLC domain and Hox-like domain common to almost all CerS. Expression of CerS2 is transiently increased during periods of active myelination, suggesting that it is important for the synthesis of myelin sphingolipids. The lack of CerS2, as shown in knockout mice, induces the autophagy and activation of the unfolded protein response (UPR). These mice showed no decrease in overall ceramide level, but levels of sphinganine were elevated. They also developed severe liver disease, but there was no observable change in the kidneys. The CerS2 gene is compact in size and is located in a chromosomal region that is replicated early in the cell cycle. CerS2 activity is regulated by sphingosine-1-phosphate (S1P) via two sphingosine-1-phosphate receptor-like residues on CerS2 that operate independently.
Background References
1. Khan SR et al. Reduced circulating sphingolipids and CERS2 activity are linked to T2D risk and impaired insulin secretion. Sci Adv. 2025 Jan
2. Wang H et al. Identifying MSMO1, ELOVL6, AACS, and CERS2 related to lipid metabolism as biomarkers of Parkinson\'s disease. Sci Rep. 2024 Jul
Western blot analysis of CerS2 on different lysates with Rabbit anti-CerS2 antibody (HA724549) at 1/2,000 dilution.
Lane 1: HepG2 (Human liver cancer cell) cell lysate
Lane 2: Huh7 (Human liver cancer cell) cell lysate
Lane 3: SH-SY5Y(Human neuroblastoma cell) cell lysate
Lane 4: Jurkat (Human T-lymphoblastic cells) cell lysate
Lysates/proteins at 20 µg/Lane.
Exposure time: 1 minute; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724549, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 45 kDa
Observed band size: 40 kDa
Application: Immunocytochemistry (IF-cell)
Species: Human
Sample: Hep G2 (Human hepatocellar carcinoma cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA724549, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724549, 1/2,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
Skeletal muscle exhibits low expression levels of CerS2 (PMID: 18165233).
Immunoprecipitation (IP)
CerS2 was immunoprecipitated in 0.2 mg Hep G2 (Human hepatocellar carcinoma cell) cell lysate with HA724549 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724549 at 1/2,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: Hep G2 cell lysate (input)
Lane 2: HA724549 IP in Hep G2 cell lysate
Lane 3: Rabbit IgG instead of HA724549 in Hep G2 cell lysate
Exposure time: 20 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary dilution: HA724549, 1/2,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Predicted band size: 45 kDa
Observed band size: 40 kDa
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