- PA5-61136 - Invitrogen Antibodies RNF121 antibody

Wang T, Hao Z, Liu C, Yuan L, Li L, Yin M, Li Q, Qi Z, Wang Z
Cell death & disease 2020 Jul 30;11(7):598

Madigan VJ, Yuziuk JA, Chiarella AM, Tyson TO, Meganck RM, Elmore ZC, Tse LV, Hathaway NA, Asokan A
PLoS pathogens 2019 Aug;15(8):e1007988

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of RNF121 in control (vector only transfected HEK293T lysate) and RNF121 over-expression lysate (Co-expressed with a C-terminal myc-DDK tag (~3.1 kDa) in mammalian HEK293T cells). Samples were probed using a RNF121 Polyclonal Antibody (Product # PA5-61136).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent staining of RNF121 in human cell line U-2 OS shows positivity in nucleus, the Golgi apparatus & vesicles. Samples were probed using a RNF121 Polyclonal Antibody (Product # PA5-61136).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical staining of RNF121 in human testis using a RNF121 Polyclonal Antibody (Product # PA5-61136) shows moderate cytoplasmic positivity in cells in seminiferous ducts.

Supportive validation

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 2 Altered LEF1 modulates the expression of circRNF121 in OA. a The expression of LEF1 was identified in OA and healthy human cartilage tissues by q-PCR. b The LEF1 expression in chondrocytes treated with IL-1beta was detected by q-PCR. c , d ChIP analysis of human chondrocytes treated with IL-1beta using specific antibodies against LEF1 was shown. e Expression of RNF121 between OA and normal groups was identified by q-PCR. f RNF121 expression in LEF1-altered expressing chondrocytes was detected by western blot. g circRNA levels were identified by q-PCR. h CircRNF121 level was positively correlated with the modified Mankin's scores. i Correlation analysis between the expression levels of LEF1 and circRNF121 was identified. j Exon2 and Exon3 region of RNF121 pre-mRNA was back- spliced and formed circRNF121. k CircRNF121 was amplified by divergent primers in cDNA but not in gDNA. GAPDH was used as a negative control. l Expression of circRNF121 was detected by q-PCR after transfection. Data were means +- SD of three independent assays (* P < 0.05).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 4 CircRNF121 functions as a sponge of miR-665 and indirectly mediates MYD88 level. a, b Three miRNAs were selected and analyzed by q-PCR. c The expression of miR-665 was negatively correlated with the modified Mankin's scores. d The correlation analysis of the expressions of circRNF121 and miR-665 was shown. e The predicted binding sites between circRNF121 and miR-665 were presented. f The dual-reporter luciferase assay confirmed that circRNF121 was the direct target of miR-665. g RNA immunoprecipitation was performed in chondrocytes transfected with miR-NC and miR-665 mimic. CircRNF121 expression was detected by using qRT-PCR. RNA levels were presented as fold enrichment in Ago2 relative to IgG immunoprecipitates. h The localization of RNF121 and miR-665 was detected by FISH assay in human chondrocytes. i The predicted binding sites between MYD88 and miR-665 were shown. j The decreased luciferase was also shown to determine the direct binding of MYD88 and miR-665. k , l MYD88 expression was determined by western blot and IF staining in treated chondrocytes. Data were means +- SD of three independent assays (* P < 0.05).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 1 Effect of RNF121 knockout (KO) and overexpression (OVX) on AAV transduction. A, Western blot of whole cell lysates from Scr and RNF121 KO cell lines. B, Luciferase reporter expression of Scr and RNF121 KO Huh7 cells following transduction with AAV1 (10,000 vg/cell), AAV2 (5,000 vg/cell), AAV6 (10,000 vg/cell), and AAV9 (20,000 vg/cell). C, Luciferase reporter expression of Scr and RNF121 KO in HEK293 and U87 cell lines following transduction with AAV2. D, Luciferase reporter signal with increasing dose of AAV2 on Scr and RNF121 KO Huh7 cells. E, Luciferase reporter signal of Scr and RNF121 KO HEK293 cells following transfection with a control plasmid or plasmid containing RNF121 cDNA. A two-tailed unpaired t test was used unless otherwise indicated, with *, p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 5 Effect of RNF121 KO on AAV capsid localization and ubiquitination. A, Representative images from confocal microscopy of Scr and RNF121 KO Huh7 cells at 12 hours post-transduction with AAV2, immunostaining for AAV2 Capsid (A20, Red), RNF121 (green), nuclei (DAPI/blue), and merged. B, Immunoprecipitation (IP) of biotinylated AAV2 capsids and immunoblotting of input and pull down material for actin, capsid, and RNF121. In vitro ubiquitination of capsids followed by immunoblotting for VP1,2,3 proteins (C) and ubiquitin (D). E, Immunoblot of RNF121 expression from catalytically dead mutants (C226A/C229A; V228A). F, AAV2-Luciferase transduction of RNF121 KO HEK293 cells following transfection of catalytically dead mutants. A two-tailed unpaired t test was used unless otherwise indicated, with *, p

PA5-61136