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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [2]
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- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
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- Product number
- PA5-29731 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Emerin Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: HeLa nucleus.
- Concentration
- 1 mg/mL
Submitted references Combined loss of LAP1B and LAP1C results in an early onset multisystemic nuclear envelopathy.
Topokaryotyping demonstrates single cell variability and stress dependent variations in nuclear envelope associated domains.
Fichtman B, Zagairy F, Biran N, Barsheshet Y, Chervinsky E, Ben Neriah Z, Shaag A, Assa M, Elpeleg O, Harel A, Spiegel R
Nature communications 2019 Feb 5;10(1):605
Nature communications 2019 Feb 5;10(1):605
Topokaryotyping demonstrates single cell variability and stress dependent variations in nuclear envelope associated domains.
Jurisic A, Robin C, Tarlykov P, Siggens L, Schoell B, Jauch A, Ekwall K, Sørensen CS, Lipinski M, Shoaib M, Ogryzko V
Nucleic acids research 2018 Dec 14;46(22):e135
Nucleic acids research 2018 Dec 14;46(22):e135
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using Emerin Polyclonal Antibody (Product # PA5-29731). Sample (30 µg of whole cell lysate). Lane A: HeLa nucleus. 12% SDS PAGE. Emerin Polyclonal Antibody (Product # PA5-29731) diluted at 1:5,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Emerin was achieved by transfecting HeLa with EMD specific siRNAs (Silencer® select Product # s225840, s4645). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the EMD knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Emerin Polyclonal Antibody (Product # PA5-29731, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant 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). Few uncharacterized bands were observed between 60-80 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using anti- Emerin Polyclonal Antibody (Product # PA5-29731) and a 30 kDa band corresponding to EMD was observed across cell lines tested along with an uncharacterized band at ~ 80 kDa. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), K-562 (Lane 2), U-87MG (Lane 3), A-431 (Lane 4) and MCF-7 (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 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).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Emerin in paraformaldehyde-fixed HeLa cells using an Emerin polyclonal antibody (Product # PA5-29731) at a 1:500 dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Emerin 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 Emerin Polyclonal Antibody (Product # PA5-29731) at 1:250 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) 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 nuclear membrane and 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 analysis of paraffin-embedded Hela xenograft, using Emerin (Product # PA5-29731) antibody at 1:1,000 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
