Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [4]
- Immunocytochemistry [3]
- Immunohistochemistry [3]
- Flow cytometry [1]
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- Product number
- PA5-79744 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- NME2 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 500 µg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of NME2 in Lane 1: rat heart tissue lysate, Lane 2: rat brain tissue lysate, Lane 3: rat liver tissue lysate, Lane 4: rat skeletal muscle tissue lysate, Lane 5: PANC cell lysate, Lane 6: HeLa cell lysate, Lane 7: SMMC cell lysate, Lane 8: U87 cell lysate, Lane 9: COLO320 cell lysate. Sample was incubated with NME2 polyclonal antibody (Product # PA5-79744).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of NME2 in, Lane 1: rat heart tissue lysates, Lane 2: rat brain tissue lysates, Lane 3: mouse heart tissue lysates, Lane 4: mouse brain tissue lysates, . Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 µg of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. The membrane was blocked with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with NME2 Polyclonal Antibody (Product # PA5-79744) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5,000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit. A specific band was detected for NME2 at approximately 17KD. The expected band size for NME2 is at 17KD.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Nucleoside diphosphate kinase B was achieved by transfecting MCF7 with Nucleoside diphosphate kinase B specific siRNAs (Silencer® select Product # S9591, S9592). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the Nucleoside diphosphate kinase B knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with NME2 Polyclonal Antibody (Product # PA5-79744, 0.5 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). 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 Nucleoside diphosphate kinase B.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot was performed using Anti-NME2 Polyclonal Antibody (Product # PA5-79744) and a 17kDa band corresponding to Nucleoside diphosphate kinase B was observed across cell lines and tissues tested . Nuclear enriched extracts (30 µg lysate) of A549 (Lane 1), HeLa (Lane 2), PC-3 (Lane 3), MCF7 (Lane 4), Hep G2 (Lane 5), K-562 (Lane 6), Jurkat (Lane 7), NIH/3T3 (Lane 8), Mouse Liver (Lane 9), Mouse Testis (Lane 10) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). 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 (0.5 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005).A dimer like pick up was observed across cell lines tested.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry analysis of NME2 in HeLa cells. Sample was incubated with NME2 polyclonal antibody (Product # PA5-79744).
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunocytochemistry analysis of NME2 using anti-NME2 antibody (Product # PA5-79744). NME2 was detected in a section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and then incubated with 2μg/mL rabbit anti-NME2 antibody (Product # PA5-79744)overnight at 4°C. DyLight®550 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Nucleoside diphosphate kinase B 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 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with NME2 Polyclonal Antibody (Product # PA5-79744) at 0.5 µg/mL 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), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Nucleus and cytoplasm localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemistry analysis of NME2 on paraffin-embedded human intestinal cancer tissue. Sample was incubated with NME2 polyclonal antibody (Product# PA5-79744).
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemistry analysis of NME2 on paraffin-embedded rat Cerebellum tissue. Sample was incubated with NME2 polyclonal antibody (Product# PA5-79744).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of NME2 in paraffin-embedded section of mouse intestine tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti-NME2 antibody (Product # PA5-79744) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometry of NME2 in HL-60 cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with NME2 Polyclonal Antibody (Product # PA5-79744) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.