Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunohistochemistry [1]
- Other assay [2]
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Validation data
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- Product number
- PA5-32081 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-PHD1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- PA5-32081 targets EGLN2 in IF, IHC (P), and WB applications and shows reactivity with Human samples. The PA5-32081 immunogen is recombinant fragment corresponding to a region within amino acids 141 and 407 of Human EGLN2. Store product as a concentrated solution. Centrifuge briefly prior to opening the vial. For short-term storage (1-2 weeks), product can be stored at 4°C. For long-term storage, aliquot and store product at -20° C or below, avioiding multiple freeze-thaw cycles.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of EGLN2 using 30 µg of HeLa lysate. Samples were loaded onto a 10% SDS-PAGE gel and probed with an EGLN2 polyclonal antibody (Product # PA5-32081) at a dilution of 1:5000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-PHD1 Polyclonal Antibody (Product # PA5-32081) and a 50 kDa band corresponding to PHD1 was observed across all the cell lines and tissues tested along with an uncharacterized band at ~55 kDa. Modified whole cell extracts (1% SDS) (30 µg lysate) of A549 (Lane 1), HeLa (Lane 2), MCF7 (Lane 3), MDA-MB-231 (Lane 4), HEL 92.1.7 (Lane 5), Caco-2 (Lane 6) and tissue extract of Mouse Brain (Lane 7) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 (1:1000 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)
- Main image
- Experimental details
- Immunohistochemical analysis of EGLN2 in paraffin-embedded lung adenocarcinoma using an EGLN2 polyclonal antibody (Product # PA5-32081) at a dilution of 1:250.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of EGLN2 in paraformaldehyde-fixed HeLa cells using an EGLN2 polyclonal antibody (Product # PA5-32081) (Green) at a 1:500 dilution. Alpha-tubulin filaments were labeled with Product # PA5-29281 (Red) at a 1:2000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of PHD1 was performed using 70% confluent log phase HeLa cells treated with Cobalt chloride. 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 BNIP3L Polyclonal Antibody (Product # PA5-32081) at 5 µg/ml dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant 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 ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing increase in Nuclear localization. Panel e represents untreated cells with Nuclear localization. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.