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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [5]
- Immunocytochemistry [1]
- Other assay [1]
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Validation data
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- Product number
- PA5-95730 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- ASM Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Positive Samples: THP-1, HepG2, U-251MG, MCF-7, Mouse brain, Mouse liver, Rat brain, Rat liver; Cellular Location: Lysosome Immunogen sequence: PARLHRIVPR LRDVFGWGNL TCPICKGLFT AINLGLKKEP NVARVGSVAI KLCNLLKIAP PAVCQSIVHL FEDDMVEVWR RSVLSPSEAC GLLLGSTCGH WDIFSSWNIS LPTVPKPPPK PPSPPAPGAP VSRILFLTDL HWDHDYLEGT DPDCADPLCC RRGSGLPPAS RPGAGYWGEY SKCDLPLRTL ESLLSGLGPA GPFDMVYWTG DIPAHDVWHQ TRQDQLRALT TVTALVRKFL GPVPVYPAVG NHESTPVNSF PPPFIEGNHS S
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.42 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
Submitted references High levels of modified ceramides are a defining feature of murine and human cancer cachexia.
Morigny P, Zuber J, Haid M, Kaltenecker D, Riols F, Lima JDC, Simoes E, Otoch JP, Schmidt SF, Herzig S, Adamski J, Seelaender M, Berriel Diaz M, Rohm M
Journal of cachexia, sarcopenia and muscle 2020 Dec;11(6):1459-1475
Journal of cachexia, sarcopenia and muscle 2020 Dec;11(6):1459-1475
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of extracts of THP-1 cells, using SMPD1 Polyclonal antibody (Product # PA5-95730) at 1:1000 dilution. THP-1 cells were treated by PMA (80 nM) for overnight. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Exposure time: 30s.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of extracts of various cell lines, using SMPD1 Polyclonal antibody (Product # PA5-95730) at 1:1000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Exposure time: 30s.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of ASM in extracts of various cell lines using ASM Polyclonal Antibody (Product # PA5-95730) at a dilution of 1:1000. A HRP Goat Anti-Rabbit IgG (H+L) secondary antibody was used at a dilution of 1:10,000. Lysates/proteins: 25 µg per lane. Blocking buffer: 3% nonfat dry milk in TBST.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of ASM in extracts of THP-1 cells using ASM Polyclonal Antibody (Product # PA5-95730) at a dilution of 1:1000. A HRP Goat Anti-Rabbit IgG (H+L) secondary antibody was used at a dilution of 1:10,000. Lysates/proteins: 25 µg per lane. Blocking buffer: 3% nonfat dry milk in TBST.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of ASM in extracts of various cell lines using ASM Polyclonal Antibody (Product # PA5-95730) at a dilution of 1:1000. A HRP Goat Anti-Rabbit IgG (H+L) secondary antibody was used at a dilution of 1:10,000. Lysates/proteins: 25 µg per lane. Blocking buffer: 3% nonfat dry milk in TBST.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry-Immunofluorescence analysis of ASM was performed in NIH-3T3 cells using ASM Polyclonal Antibody (Product # PA5-95730) at a dilution of 1:100. Blue: DAPI for nuclear staining.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 3 Expression of key enzymes involved in sphingolipid metabolism is altered in metabolic organs. ( A , B ) Liver mRNA levels of enzymes involved in ceramide metabolism in phosphate-buffered saline (PBS) (grey bars, n = 8 animals), non-cachectic NC26 (blue bars, n = 7 animals), and cachectic C26 (C26-cx, red bars, n = 6 animals) tumour-bearing mice ( A , experiment 2 ); and PBS (grey bars, n = 10 animals), pre-cachectic (C26-precx, pink bars, n = 11 animals), and cachectic (C26-cx, red bars, n = 9 animals) C26 tumour-bearing mice ( B , experiment 3 ). ( C , D ) Liver protein levels of Cers6 and Smpd1 from experiments 2 and 3 . Vinculin was used as loading control. ( E - I ) mRNA levels of enzymes involved in ceramide metabolism in epididymal white adipose tissue (eWAT) ( E , F ), GC muscle ( G , H ), and tumour ( I ). Data are mean +- SEM; statistical analyses were performed using unpaired one-way ANOVA or Kruskal-Wallis tests with Bonferroni or Dunn's post-hoc tests, respectively. Tests were two sided. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
