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- Product number
- PA5-25339 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TRAG3 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 400 µL
- Concentration
- 0.5 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
Submitted references CPEB4-Promoted Paclitaxel Resistance in Ovarian Cancer In Vitro Relies on Translational Regulation of CSAG2.
Zhang Y, Gan H, Zhao F, Ma X, Xie X, Huang R, Zhao J
Frontiers in pharmacology 2020;11:600994
Frontiers in pharmacology 2020;11:600994
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- FIGURE 3 CPEB4 binds with CSAG2 mRNAs and induces its cytoplasmic polyadenylation and translation (A,B) HEK293T cells were transfected with 5 or 10 µg plasmids expressing vector or human CPEB4 as indicated, and the cell lysates were immunoprecipitated with IgG isotype control antibody or CPEB4 antibody. The protein level of CPEB4 in whole cell lysates and IP products was analyzed by Western blotting (A) . Total RNAs bound with agarose beads were purified from immunoprecipitates and reversely transcribed to cDNA and analyzed by qRT-PCR to measure the level of CSAG2 transcript (B) . The results in each sample represent the mean value of three replicates. The enrichment value relative to IgG group is shown. Data are mean +- SEM. ** p < 0.01. (C-E) HEK293T cells were transfected with 5 or 10 µg plasmids expressing vector or human CPEB4 as indicated and cultured for 3 days. (C) Total RNAs were then extracted and the polyadenylation was measured by RNA ligation-coupled RT-PCR using ( 32 P) alpha-dATP and specific primers for CSAG2. Products were separated in a denaturing 8% polyacrylamide gel and visualized by autoradiography. *, non-adenylated RNAs; AA-, adenylated RNAs. (D) The whole cell lysates were analyzed by Western blotting for detecting the level of CSAG2. beta-Actin was used as a loading control. Shown here are representative images. (E) The mRNA of CSAG2 was analyzed by qRT-PCR. beta-Actin was used as a reference control. Data are mean +- SEM. n = 3. NS, not
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- FIGURE 4 CSAG2 is more polyadenylated and translated by CPEB4 in paclitaxel-resistant ovarian cancer cells and recurrent ovarian tumors. (A-C) Cells of SKOV3 (R), SKOV3 (S), CaOV3 (S), CaOV3 (R) were routinely cultured for 3 days. (A) Total RNAs were extracted and the polyadenylation was assessed as described before. *, non-adenylated RNAs; AA-, adenylated RNAs. (B) The protein level of CSAG2 in cell lysates was analyzed by Western blotting. beta-Actin was used as a loading control, and representative images are shown. (C) The mRNA of CSAG2 was analyzed by qRT-PCR. beta-Actin was used as a reference control. Data are mean +- SEM. n = 3. NS, not significant. (D) Representative images (left) of immunohistochemical staining of CSAG2 from matched primary and recurrent ovarian tumors treated with paclitaxel-based chemotherapy. Scale bar, 50 um. H-score (right) of CSAG2 staining is used to semi-quantify its expression levels. The black line inside the box is the median, and the lines above and below the box indicate the maximum and minimum of the H-scores. Each group contained 18 paired samples. (E) The mRNA levels of CPEB4 in matched primary and recurrent ovarian tumors (D) were analyzed by qRT-PCR. beta-Actin was used as a reference control. Each symbol represents the mean value of three replicates.
