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antibody from Invitrogen Antibodies
Targeting: SERPINH1
CBP1, CBP2, colligen, HSP47, SERPINH2
Western blotAntibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [2]
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- Product number
- PA5-76659 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SERPINH1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen and the purity is > 95% (by SDS-PAGE).
- 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 SERPINH1. Samples were incubated with SERPINH1 polyclonal antibody (Product # PA5-76659).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SERPINH1. Samples were incubated with SERPINH1 polyclonal antibody (Product # PA5-76659).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Serpin H1 was achieved by transfecting Caco-2 with Serpin H1 specific siRNAs (Silencer® select Product # s2485, s2486). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Serpin H1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with SERPINH1 Polyclonal Antibody (Product # PA5-76659, 1:1000 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 Serpin H1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-SERPINH1 Polyclonal Antibody (Product # PA5-76659) and a 47 kDa band corresponding to Serpin H1 was observed across tested cell lines with an uncharacterized band (*) at ~45 kDa. Whole cell extracts (40 µg lysate) of NIH:OVCAR-3 (Lane 1), HeLa (Lane 2), Caco-2 (Lane 3), NIH/3T3 (Lane 4), U-937 (Lane 5), Daudi (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 SERPINH1 in HeLa cells. Samples were incubated with SERPINH1 polyclonal antibody (Product # PA5-76659).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Serpin H1 was performed using 70% confluent log phase Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 5 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with SERPINH1 Polyclonal Antibody (Product # PA5-76659) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), 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 cytoplasm (endoplasmic reticulum) localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
