- 710276 - Invitrogen Antibodies TGFB2 antibody

Submitted references Dexamethasone-induced inhibition of miR-132 via methylation promotes TGF-β-driven progression of pancreatic cancer.

Abukiwan A, Nwaeburu CC, Bauer N, Zhao Z, Liu L, Gladkich J, Gross W, Benner A, Strobel O, Fellenberg J, Herr I
International journal of oncology 2019 Jan;54(1):53-64

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of TGFB2 in mouse kidney lysate (lane 1) and A549 cell lysate (lane 2) using a TGFB2 Recombinant Rabbit Polyclonal Antibody (Product # 710276) at a dilution of 2 µg/mL. Samples were detected using chemiluminescence (ECL). Results show a band at ~48kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole-cell extracts (30 µg lysate) of T98G (Lane 1), A549 (Lane 2), and NIH/3T3 (Lane 3). The blots were probed with Rabbit Anti-TGF beta-2 Recombinant Rabbit Polyclonal Antibody (Product # 710276, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 47 kDa band corresponding to TGF beta-2 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of TGFB2 in mouse kidney lysate (lane 1) and A549 cell lysate (lane 2) using a TGFB2 Recombinant Rabbit Polyclonal Antibody (Product # 710276) at a dilution of 2 µg/mL. Samples were detected using chemiluminescence (ECL). Results show a band at ~48kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of TGF beta-2 was done on U-87 MG cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with TGF beta-2 Recombinat Rabbit Polyclonal Antibody (Product # 710276, red histogram) or with rabbit isotype control (pink histogram) at 3-5 µg/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat anti-Rabbit Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 miR-132 binds to the 3'UTR of TGF-beta2 to inhibit its expression. (A) Schematic of the miR-132 binding site in the TGF-beta2 3'UTR at position 746-753 and its sequence homology to miR-132. The mutant version of the TGF-beta2 3'UTR is shown. (B) The wt and mt TGF-beta2 3'UTRs were cloned into a pLightSwitch Renilla plasmid and transfected into AsPC-1, PANC-1 and ASAN-PaCa cells in the presence or absence of 50 nM miR-132 mimics. Negative mimics served as a control. Co-transfection with Firefly luciferase (0.25 ng/ u l) served as a normalization control. At 48 h after transfection, the expression of Renilla and Firefly luciferases was detected using a FLUOstar Omega microplate reader. Renilla luciferase activities were normalized to Firefly luciferase activities. (C) PANC-1 cells were transfected with miR-132 or non-coding miRNA (NC), or mock-treated without miRNA (CO). Proteins were harvested 48 h later and analyzed by western blotting. beta-actin served as the normalization control. (D) Representative paraffin-embedded PDA tissue sections from patients with documented pre-operative intake of inhaled (n=8) or oral (n=6) GCs (+GC, n=14) and from those without GC intake (-GC, n=20) were evaluated by IHC to detect the expression of TGF-beta2. A semi-quantitative scoring system was used to evaluate expression levels based on visual determination of the percentage of positive cells. The sections were analyzed at x400 magnification, and representative images are s

710276

Antibody data