Antibody data
- Antibody Data
- Antigen structure
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- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [1]
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Validation data
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- Product number
- PA5-50678 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- hnRNP H1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- The antibody detects endogenous levels of total hnRNP H1 protein.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 2.1 mg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of HNRNPH1 was performed by loading (from left to right): LoVo and A375 cell, mouse brain tissue, HepG2, A431 and Hela cell lysates (40µg) on to a 8% SDS-PAGE gel. Proteins were transferred to a membrane and the membrane was probed with a HNRNPH1 antibody (Product # PA5-50678) at a 1/350 dilution for 40 seconds, followed by a secondary antibody at 1/8000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of hnRNP H1 was achieved by transfecting Hep G2 with hnRNP H1 specific siRNAs (Silencer® select Product # s6728, s6729). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1%SDS) from the knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with hnRNP H1 Polyclonal Antibody (Product # PA5-50678, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/mL, 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 hnRNP H1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-hnRNP H1 Polyclonal Antibody, (Product # PA5-50678) and a 50 kDa band corresponding to hnRNP H1 was observed in the cell lines tested. Modified whole cell extracts (1%SDS) (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), BeWo (Lane 3), PANC-1 (Lane 4) and Jurkat (Lane 5) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 HNRNPH1 was performed using 70% confluent log phase HeLa 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 1 hour at room temperature. The cells were labeled with hnRNP H1 Polyclonal Antibody (Product # PA5-50678) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790) at a dilution of 1:2000 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). Panel d represents the merged image showing staining in nucleus. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.