Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [4]
- Immunohistochemistry [2]
- Flow cytometry [1]
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Validation data
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- Product number
- MA5-32794 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- alpha Actinin 4 Recombinant Rabbit Monoclonal Antibody (JU20-23)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- JU20-23
- 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 alpha Actinin 4 in different lysates using a Monoclonal antibody (Product #MA5-32794) at a dilution of 1:500. Positive control: Lane 1: Hela, Lane 2: PC-12, Lane 3: NIH-3T3, Lane 4: Rat liver tissue, Lane 5: A431, Lane 6: HepG2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of alpha Actinin 4 in different lysates using a Monoclonal antibody (Product #MA5-32794) at a dilution of 1:500. Positive control: Lane 1: Hela, Lane 2: PC-12, Lane 3: NIH-3T3, Lane 4: Rat liver tissue, Lane 5: A431, Lane 6: HepG2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed using Anti-alpha Actinin 4 Recombinant Rabbit Monoclonal Antibody (JU20-23) (Product # MA5-32794) and a ~110 kDa band corresponding to Alpha-actinin-4 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), A-431 (Lane 3), IMR-90 (Lane 4), PANC-1 (Lane 5), DU 145 (Lane 6), LNCaP (Lane 7), PC-12 (Lane 8) and NIH/3T3 (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The Blot was probed with the primary antibody (1:2000 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). https://doi.org/10.1016/j.bbrc.2019.12.084
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of alpha Actinin 4 in A549 cells using a alpha Actinin 4 Monoclonal antibody (Product # MA5-32794) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of alpha Actinin 4 in Hela cells using a alpha Actinin 4 Monoclonal antibody (Product # MA5-32794) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of alpha Actinin 4 in LOVO cells using a alpha Actinin 4 Monoclonal antibody (Product # MA5-32794) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Alpha-actinin-4 was performed using 70% confluent log phase DU 145 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 alpha Actinin 4 Recombinant Rabbit Monoclonal Antibody (JU20-23) (Product # MA5-32794) 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 Cytoplasmic 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 alpha Actinin 4 of paraffin-embedded Human colon cancer tissue using a alpha-Actinin-4 Monoclonal antibody (Product #MA5-32794). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of alpha Actinin 4 of paraffin-embedded Mouse colon tissue using a alpha-Actinin-4 Monoclonal antibody (Product #MA5-32794). Counter stained with hematoxylin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometric analysis of alpha Actinin 4 in A549 cells using a alpha Actinin 4 Monoclonal Antibody (Product # MA5-32794) at a dilution of 1:100, as seen in red compared with an unlabelled control (cells without incubation with primary antibody; black).