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LearnMA1-16846
antibody from Invitrogen Antibodies
Targeting: DIABLO
DFNA64, DIABLO-S, FLJ10537, FLJ25049, SMAC
Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
- References [0]
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- Validations
- Western blot [4]
- Immunocytochemistry [1]
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- Product number
- MA1-16846 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- DIABLO Monoclonal Antibody (78-1-118)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- Suggested positive control: 293 whole cell lysate.
- Reactivity
- Human, Mouse, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 78-1-118
- Vial size
- 200 µL
- Concentration
- Conc. Not Determined
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Smac on HeLa whole cell extract lysates.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of DIABLO in HeLa whole cell extract lysates. Sample was incubated in DIABLO monoclonal antibody (Product # MA1-16846).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of DIABLO was achieved by transfecting HeLa with DIABLO specific siRNAs (Silencer® select Product # S32187, S32188). Western blot analysis (Fig. a) was performed using Whole cell extracts from the DIABLO knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with DIABLO Monoclonal Antibody (78-1-118), (Product # MA1-16846, 1:500 dilution ) 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 DIABLO.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-DIABLO Monoclonal Antibody (78-1-118), (Product # MA1-16846) and a 21kDa band corresponding to DIABLO was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), MCF-7 (Lane 2) and Hep G2 (Lane 3) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) 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-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000 dilution) using the iBright FL1000 (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
- Immunofluorescence analysis of DIABLO was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with DIABLO Monoclonal Antibody (78-1-118) (Product # MA1-16846) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing Mitochondrial localization. Panel e represents control HeLa cells with no primary antibody to assess background. The images were captured at 60X magnification.
