- PA5-110105 - Invitrogen Antibodies PHIP antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of PHIP in HeLa, Jurkat. Samples were incubated in polyclonal PHIP antibody (Product # PA5-110105) using a dilution of 1:1000, followed by HRP Goat Anti-Rabbit IgG (H+L) at a dilution of 1:10000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry analysis of PHIP in MCF-7. Samples were incubated in polyclonal PHIP antibody (Product # PA5-110105) using a dilution of 1:100.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of PH-interacting protein (PHIP) was performed using 70% confluent log phase A-431 and U-87 MG 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 10 minutes at room temperature. The cells were labeled with PHIP Polyclonal Antibody (Product # PA5-110105) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (1:2500), 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 for PHIP in A-431 cells and no signal in U-87 MG (Panel e), which is reported low expressing for the protein. Panel f represents control A-431 cells with no primary antibody to assess background. The images were captured at 40X magnification with CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7C1115LZR).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of PH-interacting protein (PHIP) was achieved by transfecting SH-SY5Y cells with PHIP specific siRNA (Silencer® select Product # s30010, s30012). Immunofluorescence analysis was performed on untransfected SH-SY5Y cells (panel a-d), transfected with non-specific scrambled siRNA (panels e-h) and transfected with PHIP specific siRNA (panel i-l). Cells were fixed, permeabilized, and labelled with PHIP Polyclonal Antibody (Product # PA5-110105, 1:100) followed by Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2500 dilution). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (Red) staining. Reduction of specific signal was observed upon siRNA mediated knockdown (panel i,l) confirming specificity of the antibody to PHIP (Green). The images were captured at 60X magnification in EVOS™ M7000 Imaging System (Product # AMF7000).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of PHIP in paraffin-embedded human kidney cancer tissue. Samples were incubated in polyclonal PHIP antibody (Product # PA5-110105) using a dilution of 1:100.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of PHIP in paraffin-embedded well-differentiated human squamous skin carcinoma tissue. Samples were incubated in polyclonal PHIP antibody (Product # PA5-110105) using a dilution of 1:100.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of PHIP in paraffin-embedded mouse tumor tissue. Samples were incubated in polyclonal PHIP antibody (Product # PA5-110105) using a dilution of 1:100.

PA5-110105