Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [1]
- Flow cytometry [1]
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Validation data
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- Product number
- PA1-16525 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PHD2 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody does not react with mouse samples.
- Concentration
- 1 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of human PHD2, using Product # PA1-16525. Samples: Recombinant FLAG-His-PHD1, PHD2 and PHD3 (10 ng/lane), HeLa whole cell lysate and MEFs.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of PHD2 was achieved by transfecting HeLa with PHD2 specific validated siRNAs (Silencer® select Product # s29231). Western blot analysis (Fig. a) was performed using whole cell extracts from the PHD2 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with PHD2 Polyclonal Antibody (Product # PA1-16525, 1 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to PHD2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of HEK 293 (Lane 1), COS-7 (Lane 2) and U-87 MG (Lane 3). The blot was probed with Anti-PHD2 Polyclonal Antibody (Product # PA1-16525, 1 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 46 kDa band corresponding to PHD2 was observed across the cell lines tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of PHD2 in 10 ng/lane recombinant FLAG-His-PHD1, PHD2 and PHD3 , HeLa whole cell lysate and MEFs. Samples were incubated in PHD2 polyclonal antibody (Product # PA1-16525.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of PHD2 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 1% BSA for 1 hour at room temperature. The cells were labeled with PHD2 Polyclonal Antibody (Product # PA1-16525) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 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). Panel d represents the merged image showing nucleus and cytoplasmic 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)
- Main image
- Experimental details
- Flow cytometry of PHD2 in Jurkat cells (fixed, permeabilized). Samples were incubated in PHD2 polyclonal (Product # PA1-16525) using a dilution of 3.0 µg/mL (in a 150 µL reaction).