Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-27487 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PRDX1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, H1299, HeLa, HepG2, Molt-4, Raji, NIH-3T3. Predicted reactivity: Mouse (95%), Rat (98%), Zebrafish (80%), Dog (97%), Chicken (86%), Rhesus Monkey (98%), Bovine (95%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.22 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 using PRDX1 Polyclonal Antibody (Product # PA5-27487). Sample (30 µg of whole cell lysate). Lane A:NIH-3T3 12% SDS PAGE. PRDX1 Polyclonal Antibody (Product # PA5-27487) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using PRDX1 Polyclonal Antibody (Product # PA5-27487). Sample (30 µg of whole cell lysate). Lane A: 293T. 12% SDS PAGE. PRDX1 Polyclonal Antibody (Product # PA5-27487) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of PRDX1 was achieved by transfecting HeLa with PRDX1 specific siRNAs (Silencer® select Product # s10007, s224164). Western blot analysis (Fig. A) was performed using Whole cell extracts from the PRDX1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with PRDX1 Polyclonal Antibody (Product # PA5-27487, 1:1000) 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 PRDX1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-PRDX1 Polyclonal Antibody (Product # PA5-27487) and a 22kDa band corresponding to PRDX1 was observed across all the tested cell lines, and was upregulated upon Hydrogen peroxide treatment. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HeLa (Hydrogen peroxide, 0.5mM, 2 hrs) (Lane 2), MCF7 (Lane 3), MCF7 (Hydrogen peroxide, 0.5mM, 2 hrs) (Lane 4), HCT 116 (Lane 5), A549 (Lane 6) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). 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) 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
- Immunofluorescent analysis of PRX I in methanol-fixed HeLa cells using a PRX I polyclonal antibody (Product # PA5-27487) at a 1:200 dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of PRDX1 was performed using 70% confluent log phase HeLa treated with Hydrogen peroxide (0.5mM, 2 hrs). 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 PRDX1 Polyclonal Antibody (Product # PA5-27487) at 1:200 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), (1:2000), 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 cytoplasmic localization. Panel e represents control cells showing low levels of expression of PRDX1. Panel f 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
- PRDX1 Polyclonal Antibody detects PRX I protein at cytosol on mouse lung by immunohistochemical analysis. Sample: Paraffin-embedded mouse lung. PRDX1 Polyclonal Antibody (Product # PA5-27487) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.