Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [3]
- Immunohistochemistry [4]
- Flow cytometry [1]
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Validation data
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- Product number
- MA5-32080 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- NOTCH1 Recombinant Rabbit Monoclonal Antibody (SJ205)
- 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
- SJ205
- 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 NOTCH1 in HepG2 cell lysates using a NOTCH1 Monoclonal antibody (Product # MA5-32080) at a dilution of 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockout of NOTCH1 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR682588_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of NOTCH1 was performed by loading 30 µg of MDA-MB-231 wild type (Lane 1) and MDA-MB-231 NOTCH1 KO (Lane 2) whole cell extracts. The samples were electrophoresed using using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-NOTCH1 Recombinant Rabbit Monoclonal Antibody (SJ205)(Product # MA5-32080) using 1:1,000 dilution and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to NOTCH1. Uncharacterized bands were observed between 31-41 kDa in HeLa wild type sample and ~45 kDa in both the samples.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-NOTCH1 Recombinant Rabbit Monoclonal Antibody (SJ205) (Product # MA5-32080). A 300 kDa and a 100 kDa band corresponding to NOTCH1 was observed across MCF 10A, MDA-MB-231, Molt-4, Mouse Brain and Rat Brain. Whole cell extracts (40 µg lysate) of MCF 10A (Lane 1), MDA-MB-231 (Lane 2), MCF7 (Lane 3), MOLT-4 (Lane 4), Jurkat (Lane 5), Mouse Skeletal Muscle (Lane 6), Rat Skeletal Muscle (Lane 7), Mouse Brain (Lane 8) and Rat Brain (Lane 9) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). 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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of NOTCH1 in Hela cells using a NOTCH1 Monoclonal antibody (Product # MA5-32080) 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 NOTCH1 in HepG2 cells using a NOTCH1 Monoclonal antibody (Product # MA5-32080) 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 NOTCH1 in NIH/3T3 cells using a NOTCH1 Monoclonal antibody (Product # MA5-32080) 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 NOTCH1 of paraffin-embedded Human lung tissue using a NOTCH1 Monoclonal antibody (Product #MA5-32080). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of NOTCH1 of paraffin-embedded Human spleen tissue using a NOTCH1 Monoclonal antibody (Product #MA5-32080). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of NOTCH1 of paraffin-embedded Mouse spleen tissue using a NOTCH1 Monoclonal antibody (Product #MA5-32080). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of NOTCH1 of paraffin-embedded Mouse lung tissue using a NOTCH1 Monoclonal antibody (Product #MA5-32080). Counter stained with hematoxylin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometric analysis of NOTCH1 in Hela cells using a NOTCH1 Monoclonal Antibody (Product # MA5-32080) at a dilution of 1:50, as seen in blue compared with an unlabelled control (cells without incubation with primary antibody; red). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.