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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [4]
- Immunocytochemistry [2]
- Immunoprecipitation [1]
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- Product number
- MA5-24875 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- LSD1 Monoclonal Antibody (1E5)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- The antibody was affinity-purified from mouse ascites by affinity-chromatography using epitope-specific immunogen and the purity is > 95% (by SDS-PAGE).
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 1.00E+05
- Vial size
- 100 µL
- 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 KDM1A in various cell lysate samples using KDM1A polyclonal antibody (Product # MA5-24875) at a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of LSD1 in HeLa, HCT116 and COS7 cell lysates. Samples were incubated with LSD1 monoclonal antibody (Product # MA5-24875) at a dilution of 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-LSD1 Monoclonal Antibody (1E5) (Product # MA5-24875) and a 110 kDa band corresponding to LSD1 was observed across tested cell lines. Nuclear enriched extracts (40 µg lysate) of K-562 (Lane 1), MCF7 (Lane 2), THP-1 (Lane 3), HeLa (Lane 4) 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-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:20000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of LSD1 was achieved by transfecting MCF7 with LSD1 specific siRNAs (Silencer® select Product # s617, s619). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the LSD1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with LSD1 Monoclonal Antibody (1E5) (Product # MA5-24875, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:20000 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 LSD1 along with an uncharacterized band(*) at ~60 kDa.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of LSD1 was performed using 70% confluent log phase K-562 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 1 hour at room temperature. The cells were labeled with LSD1 Monoclonal Antibody (1E5) (Product # MA5-24875) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (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 predominantly nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of LSD1 in HeLa cells. Samples were treated with 4% paraformaldehyde and incubated with LSD1 monoclonal antibody (Product # MA5-24875) at a dilution of 1:100.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunoprecipitation of LSD1 in HeLa cells. Samples were incubated with LSD1 monoclonal antibody (Product # MA5-24875).
