Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
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Validation data
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- Product number
- PA5-30125 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PPA1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: NT2D1, IMR32, U87-MG, MCF-7. Predicted reactivity: Mouse (92%), Rat (90%), Bovine (93%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.95 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 PPA1 Polyclonal Antibody (Product # PA5-30125). Sample (30 µg of whole cell lysate). Lane A: IMR32. Lane B: U87-MG. 12% SDS PAGE. PPA1 Polyclonal Antibody (Product # PA5-30125) diluted at 1:10,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- KD of PPA1 was achieved by transfecting U-87 MG with PPA1 specific siRNAs (Silencer® select Product # s10878, s10877). Western blot analysis (Fig. a) was performed using whole cell extracts from the PPA1 KD cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with PPA1 Polyclonal Antibody (Product # PA5-30125, 1:10000 Dilution) and Goat Anti Rabbit (H+L) Superclonal™ Secondary Antibody, HRP conjugate (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 PPA1. .
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-PPA1 Rabbit Polyclonal Antibody (Product # PA5-30125) and 32 kDa band corresponding to PPA1 was observed across the samples tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), U-87 MG (Lane 2), HT-29 (Lane 3),HCT 116 (Lane 4), SW-480 (Lane 5),MCF-7 (Lane 6) and tissue extract of Mouse Small Intestine (Lane 7) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris 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 (1:10000 Dilution) and detected by chemiluminescence anti Rabbit IgG Secondary Antibody, HRP conjugate (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).