PA1-22860
antibody from Invitrogen Antibodies
Targeting: SERPINA1
A1A, A1AT, AAT, alpha-1-antitrypsin, alpha1AT, PI, PI1
Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA1-22860 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- alpha-1 Antitrypsin Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Other
- Description
- Positive control: tonsil.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 500 µL
- Storage
- 4° C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of alpha-1 Antitrypsin was achieved by transfecting Hep G2 with alpha-1 Antitrypsin specific siRNAs (Silencer® select Product # S10459, S10458). Western blot analysis (Fig. a) was performed using Membrane enriched extracts from the alpha-1 Antitrypsin knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with alpha-1 Antitrypsin Polyclonal Antibody (Product # PA1-22860, 1:250 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (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 alpha-1 Antitrypsin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-alpha-1 Antitrypsin Polyclonal Antibody(Product # PA1-22860) and a 55kDa band corresponding to alpha-1 Antitrypsin was observed across cell lines tested. Membrane enriched extracts (30 µg lysate) of Hep G2 (Lane 1) and HT-29 (Lane 2) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). 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
- Immunofluorescence analysis of alpha-1 Antitrypsin was performed using 70% confluent log phase Hep G2 cells. 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 alpha-1 Antitrypsin Polyclonal Antibody (Product # PA1-22860) at 1:2000 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), (1:2000 dilution) 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 dilution). Panel d represents the merged image showing cytoplasmic (Endoplasmic reticulum) 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
- Immunohistochemical (Paraffin) analysis of human tonsil tissue using (Product # PA1-22860) alpha-1 Antitrypsin Polyclonal Antibody.