Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [6]
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Validation data
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- Product number
- PA1-16831 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- NUMB Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- The target sequence has 90% sequence homology with rat.
- Reactivity
- Human, Mouse, Chicken/Avian
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 3.5 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
- Detection of Numb isoforms 1 and 2 in A431 whole cell lysate (20 µg) using 0.5 µg/mL (Product # PA1-16831). ECL detection: 30 seconds.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of NUMB in 20 µg A431 whole cell lysate. Samples were incubated in NUMB polyclonal antibody (Product # PA1-16831 using a dilution of 0.5 µg/mL. ECL detection: 30 seconds.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of NUMB in 20 µg A431 whole cell lysate. Samples were incubated in NUMB polyclonal antibody (Product # PA1-16831 using a dilution of 0.5 µg/mL. ECL detection: 30 seconds.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of NUMB in 0.5 mg/mL A431 lysate. Samples were incubated in NUMB polyclonal antibody (Product # PA1-16831). This experiment was performed under reducing conditions using the 12-230 kDa separation system.
- 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 # PA1-16831) 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 isoforms.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-NUMB Polyclonal Antibody (Product # PA1-16831) on whole cell extracts (30 µg lysate) of U-87 MG (Lane 1), PC-3 (Lane 2), DU 145 (Lane 3), A549 (Lane 4), SH-SY5Y (Lane 5), HeLa (Lane 6) and Mouse lung (Lane 7) and numb major isoform 65, 72 kDa was observed across cell lines except A549. 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 (1:2000 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).