- PA5-29004 - Invitrogen Antibodies KHK antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Ketohexokinase using 30 µg of HeLa lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a Ketohexokinase polyclonal antibody (Product # PA5-29004) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using KHK Polyclonal Antibody (Product # PA5-29004). Sample (20 µg of whole cell lysate). Lane A: mouse Liver. 12% SDS PAGE. KHK Polyclonal Antibody (Product # PA5-29004) diluted at 1:10,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using KHK Polyclonal Antibody (Product # PA5-29004). Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with KHK Polyclonal Antibody (Product # PA5-29004) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of Ketohexokinase was achieved by transfecting Hep G2 with Ketohexokinase specific siRNAs (Silencer® select Product # S7828, S7830). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Ketohexokinase knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with KHK Polyclonal Antibody (Product # PA5-29004, 1:5000 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 Ketohexokinase.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-KHK Polyclonal Antibody (Product # PA5-29004) and a 32 kDa and corresponding to Ketohexokinase was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of MCF7 (Lane 1) and Hep G2 (Lane 2) as seen in Fig (a). Tissue extracts of Mouse Lung (Lane 1), Rat Lung (Lane 2), Mouse Brain (Lane 3), Rat Brain (Lane 4), Mouse Liver (Lane 5), Rat Liver (Lane 6), Mouse Kidney (Lane 7), Rat Kidney (Lane 8) and Mouse Skeletal Muscle (Lane 9) as seen in Fig (b) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 (1:5000 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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of KHK was performed using 70% confluent log phase SH-SY5Y 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 1 hour at room temperature. The cells were labeled with KHK Polyclonal Antibody (Product # PA5-29004) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then 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 dilution). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in EVOS™ M7000 Imaging System (Product # AMF7000).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry (Paraffin) analysis of KHK was performed in paraffin-embedded mouse kidney tissue using KHK Polyclonal Antibody (Product # PA5-29004) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry (Paraffin) analysis of KHK was performed in paraffin-embedded mouse liver tissue using KHK Polyclonal Antibody (Product # PA5-29004) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of paraffin-embedded A549 xenograft, using Ketohexokinase (Product # PA5-29004) antibody at 1:500 dilution. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: KHK Polyclonal Antibody detects Ketohexokinase protein at cytoplasm by immunohistochemical analysis. Sample: Paraffin-embedded mouse liver. Ketohexokinase stained by KHK Polyclonal Antibody (Product # PA5-29004) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

PA5-29004