Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [6]
- Immunocytochemistry [2]
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Validation data
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- Product number
- PA1-32452 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- NUMB Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human, Mouse, Rat, Chicken/Avian
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 2 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 of NUMB in A431 whole cell lysate (20 µg) using a NUMB polyclonal antibody (Product # PA1-32452) at a dilution of 0.5 µg/mL followed by ECL detection for 30 seconds. Results show detection of NUMB isoforms 1 and 2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of NUMB was performed by loading 20 µg of A431 cell lysates. Proteins were transferred to a membrane and probed with a NUMB Polyclonal Antibody (Product # PA1-32452) at a dilution of 0.5 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of NUMB was performed by loading 20 µg of A431 cell lysates. Proteins were transferred to a membrane and probed with a NUMB Polyclonal Antibody (Product # PA1-32452) at a dilution of 0.5 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of NUMB was performed by loading A431 cell lysate. Proteins were transferred to a membrane and probed with a NUMB Polyclonal Antibody (Product # PA1-32452).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of NUMB was achieved by transfecting U-87 MG cells with NUMB specific siRNAs (Silencer® select Product # s16466, Product # s16465). Western blot analysis (Fig. a) was performed using whole cell extracts from the NUMB knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed using NUMB Polyclonal Antibody (Product # PA1-32452) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP conjugate (Product # A27036, 0.25 ug/ml, 1:4000 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 NUMB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-NUMB Polyclonal Antibody (Product # PA1-32452) on Whole cell extracts (30 µg lysate) of U-87 MG (Lane 1), PC-3 (Lane 2), DU 145 (Lane 3), SH-SY5Y (Lane 4), HeLa (Lane 5), Mouse lung (Lane 6) and Mouse brain (Lane 7) and NUMB major isoforms 65, 72 kDa were observed across cell lines tested except SH-SY5Y and HeLa. 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 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
- Immunocytochemistry-Immunofluorescence analysis of NUMB in HepG2 cells using NUMB Polyclonal Antibody (Product # PA1-32452) (Green) at a dilution of 1:500. Red : Tubulin. Blue: DAPI.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of NUMB was performed using 70% confluent log phase U-87 MG 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 NUMB Polyclonal Antibody (Product # PA1-32452) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 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). 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.