Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Flow cytometry [2]
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Validation data
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- Product number
- MA5-24064 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IDE Monoclonal Antibody (334501)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- Reconstitute at 0.5 mg/mL in sterile PBS.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 334501
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Insulin-degrading enzyme was achieved by transfecting HeLa with Insulin-degrading enzyme specific siRNAs (Silencer® select Product # s7116, s7118). Western Blot analysis (Fig. a) was performed using Whole cell extracts from the Insulin-degrading enzyme knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with IDE Monoclonal Antibody (334501) (Product # MA5-24064, 1 µg/mL ) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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 Insulin-degrading enzyme.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed using Anti-IDE Monoclonal Antibody (334501) (Product # MA5-24064) and a 117 kDa band corresponding to Insulin-degrading enzyme was observed across cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), K-562 (Lane 3), A-431 (Lane 4), Caco-2 (Lane 5), U-937 (Lane 6) and C2C12 (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometric analysis of HeLa cells were stained with mouse Anti-human Insulysin/IDE Monoclonal Antibody (Product # MA5-24064) or isotype control antibodyopen histogram), followed by Allophycocyanin-conjugated Anti-mouse IgG F (ab')2 Secondary Antibody. To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry of IDE in HeLa cells. Samples were incubated in IDE monoclonal antibody (Product # MA5-24064) or isotype control antibody followed by Allophycocyanin-conjugated Anti-Mouse IgG F(ab)2 Secondary Antibody. To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.