- PA5-96893 - Invitrogen Antibodies USP26 antibody

Li G, Qi HW, Dong HG, Bai P, Sun M, Liu HY
Cancer cell international 2020;20:355

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of extracts of Mouse testis, using USP26 Polyclonal antibody (Product # PA5-96893) 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: 1s.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of USP26 in extracts of various cell lines using USP26 Polyclonal Antibody (Product # PA5-96893) 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)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of USP26 was performed in A549 cells using USP26 Polyclonal Antibody (Product # PA5-96893).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 2 Differential expression of Snail1 and USP26 protein and miR-203a-5p in normal esophagus and EC cell lines. a Representative blots from three experiments showing steady state expression of Snail1 and USP26 in the normal esophageal cell line HET-2A and the EC cell lines Kyse150 and TE-1. GAPDH. Loading control. b Relative expression of miR-203-5p in the three cell lines determined by qRT-PCR. Shown are scattered plot, mean +- SD (n = 06). ****P < 0.0001

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 4 Modulating miR-203 expression in EC cell lines impact Snail1 protein stability. a Representative blots from three experiments showing steady expression of Snail1, USP26 and E-cadherin in the normal esophageal cell line HET-1A and the EC cell lines Kyse150 and TE-1 transfected with either control or MIR203A/5P mimic. GAPDH. Loading control. b TE-1 cells transfected with control or MIR203A/5P mimic were treated with 50 uM cycloheximide (CHX) for up to 8 h and then lysates were probed for Snail1 and GAPDH protein expression. Shown are representative blots from three experiments. c Blots in b were analyzed using NIH ImageJ algorithm and half-life of Snail1 protein calculated

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 6 Suppression of miR-203-mediated regulation of USP26 is required for metastatic progression of EC cells. Xenograft model of experimental metastasis was established by lateral tail vein injection of parental Kyse150 cells or Kyse150 cells stably overexpressing USP26 into BALB/c nude mice (n = 10 and 5 mice, respectively). The mice injected with parental Kyse150 cells were randomly divided into two groups - control mimic (n = 05) ( left panels ), and MIR203A/5P mimic (n = 05) ( middle panels ), where the mice injected with Kyse150 cells overexpressing USP26 received MIR203A/5P mimic ( right panels ). The incidence of metastasis was measured by luciferin injection and bioluminescence imaging. Shown are representative animals from each group after 6 weeks ( a ). Mice were euthanized at the end of 6 weeks and the lungs from each group of experimental animals were surgically excised, fixed overnight in 10% buffered formalin. Representative images of hematoxylin and eosin (H and E) staining of the lungs are shown ( b ). Black arrows indicate micrometastasis. Scale bar, 100 um. c Representative images of IHC staining for USP26 protein in each experimental group. Scale bar, 30 um

PA5-96893