Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [3]
- Immunohistochemistry [1]
- Flow cytometry [1]
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Validation data
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- Product number
- MA5-32364 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- ATF4 Recombinant Rabbit Monoclonal Antibody (SD20-92)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- 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
- SD20-92
- 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
- CRISPR-Cas9 mediated genome editing ofATF4 (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR988840_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of ATF4 was performed by loading 30 µg of HeLa Wild Type (Lane 1), treated HeLa Wild type (Lane 2), HeLa Cas9 (Lane 3), treated HeLa Cas9 (Lane 4), HeLa Cas9 cells transduced with ATF4 Lentiviral sgRNA (Lane 5) and treated HeLa Cas9 cells transduced with ATF4 and Lentiviral sgRNA (Lane 6) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with ATF4 Recombinant Rabbit Monoclonal Antibody (SD20-92) (Product # MA5-32364) using 1:1,000 dilution and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036 1:5,000 dilution). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). A reduced signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific toATF4. Uncharacterized band was observed in all the samples at ~56 kDa. Treatment used was 2 µg/mL tunicamycin for 8 hrs.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of ATF4 in different lysates using a Monoclonal antibody (Product #MA5-32364) at a dilution of 1:1,000. Positive control: Lane 1: Hela, Lane 2: PC-12.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-ATF4 Recombinant Rabbit Monoclonal Antibody (SD20-92) (Product # MA5-32364) and a 48 kDa band corresponding to ATF4 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of MCF7 (Lane 1), MCF7 treated with 2 µM Thapsigargin for 8 hours (Lane 2), SH-SY5Y (Lane 3), SH-SY5Y treated with 1.5 µM Thapsigargin for 24 hours (Lane 4), A-375 (Lane 5), A-375 treated with 7.5 µM Thapsigargin for 24 hours (Lane 6), HeLa (Lane 7) and HeLa treated with 2 µg/ml Tunicamycin for 8 hours (Lane 8) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Uncharacterized bands (*) at 20 kDa and 58 kDa were observed.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of ATF4 in N2A cells using a ATF4 Monoclonal antibody (Product # MA5-32364) 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 ATF4 in MCF-7 cells using a ATF4 Monoclonal antibody (Product # MA5-32364) 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 ATF4 in PC-12 cells using a ATF4 Monoclonal antibody (Product # MA5-32364) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of ATF4 of paraffin-embedded Human breast carcinoma tissue using a ATF4 Monoclonal antibody (Product #MA5-32364). Counter stained with hematoxylin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometric analysis of ATF4 in Hela cells using a ATF4 Monoclonal Antibody (Product # MA5-32364) at a dilution of 1:50, as seen in red compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.