Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [4]
- Immunohistochemistry [2]
- Flow cytometry [1]
- Chromatin Immunoprecipitation [1]
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Validation data
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- Product number
- MA5-32240 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TCF7L2 Recombinant Rabbit Monoclonal Antibody (SC06-90)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- SC06-90
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TCF7L2 in Jurkat cell lysates using a TCF7L2 Monoclonal antibody (Product # MA5-32240) at a dilution of 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Transcription factor 7-like 2 was achieved by transfecting HCT 116 with Transcription factor 7-like 2 specific siRNAs (Silencer® select Product # s13881, s13882). Western Blot analysis (Fig. a) was performed using Modified whole cell extracts (1% SDS) from the Transcription factor 7-like 2 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with TCF7L2 Recombinant Rabbit Monoclonal Antibody (SC06-90) (Product # MA5-32240, 1:500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this Western Blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Transcription factor 7-like 2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed using Anti-TCF7L2 Recombinant Rabbit Monoclonal Antibody (SC06-90) (Product # MA5-32240) and a 55 kDa band corresponding to Transcription factor 7-like 2 was observed across the cell lines tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of HCT 116 (Lane 1), HeLa (Lane 2), Hep G2 (Lane 3), SW480 (Lane 4) and Caco-2 (Lane 5) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:500 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of TCF7L2 in Hela cells using a TCF7L2 Monoclonal antibody (Product # MA5-32240) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of TCF7L2 in SW480 cells using a TCF7L2 Monoclonal antibody (Product # MA5-32240) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of TCF7L2 in D3 cells using a TCF7L2 Monoclonal antibody (Product # MA5-32240) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Transcription factor 7-like 2 was performed using 70% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with TCF7L2 Recombinant Rabbit Monoclonal Antibody (SC06-90) (Product # MA5-32240) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7C1115LZR).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of TCF7L2 of paraffin-embedded Human kidney tissue using a TCF7L2 Monoclonal antibody (Product #MA5-32240). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of TCF7L2 of paraffin-embedded Human breast carcinoma tissue using a TCF7L2 Monoclonal antibody (Product #MA5-32240). Counter stained with hematoxylin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometric analysis of TCF7L2 in Jurkat cells using a TCF7L2 Monoclonal Antibody (Product # MA5-32240) at a dilution of 1:50, as seen in red compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- ChIP was performed using TCF7L2 Monoclonal Antibody (Product # MA5-32240, 5 µg) on sheared chromatin from HCT116 cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to LGR5_proximal (PR), BCL3_proximal (PR), ANKARD5_distal (PR),c-myc_WRE, c-myc (-290 kb UP) (TSS), ZNF180_3’ exon as active binding regions and CD70_ down (TSS) and SAT alpha as inactive binding regions. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method. PR-promoter; TSS- Transcription start site.