NUMB Recombinant Rabbit Monoclonal Antibody [PSH25-86]
cat.: HA751998
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Rat, Green monkey |
| Applications: | WB, FC(Intra), IP |
| Clonality: | Monoclonal |
| Clone number: | PSH25-86 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4). |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 71 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human NUMB aa 301-650. |
| Positive control: | U-87 MG (Human glioblastoma cell) cell lysates, SK-OV-3 (Human ovarian cancer cell) cell lysate, COS-1 (African green monkey kidney fibroblast) cell lysate, PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate. |
| Subcellular location: | Cell membrane, Endosome membrane. |
| Recommended Dilutions:
WB FC(Intra) IP |
1:10,000 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P49757 Human | Q2LC84 Rat |
| Alternative names: | c14_5527 C14orf41 FLJ31314 h Numb h-Numb Numb Numb homolog (Drosophila) Numb homolog Numb protein homolog NUMB_HUMAN Protein numb homolog Protein S171 S171 |
Images
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Fig1:
Western blot analysis of NUMB on U-87 MG (Human glioblastoma cell) cell lysate with Rabbit anti-NUMB antibody (HA751998) at 1/10,000 dilution.
Lysates/proteins at 15 µg/Lane. Exposure time: 59 seconds; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA751998, 1/10,000 in primary antibody dilution buffer (K1803), overnight at 4 °C Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 71 kDa Observed band size: 72/71/66/65 kDa |
|
Fig2:
Western blot analysis of NUMB on different lysates with Rabbit anti-NUMB antibody (HA751998) at 1/10,000 dilution.
Lane 1: SK-OV-3 (Human ovarian cancer cell) cell lysate Lane 2: SK-BR-3 (Human breast cancer cell) cell lysate Lane 3: COS-1 (African green monkey kidney fibroblast) cell lysate Lane 4: PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate Lysates/proteins at 15 µg/Lane. Exposure time: 3 minutes; ECL: K1801 Negative control: SK-BR-3 (PMID: 20443831). Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA751998, 1/10,000 in primary antibody dilution buffer (K1803), overnight at 4 °C Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 71 kDa Observed band size: 72/71/66/65 kDa |
|
Fig3:
Application: Flow Cytometry (Intra) Species: Rat Sample: PC-12 (Rat pheochromocytoma cells (undifferentiated)) Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature. Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature. Antibody dilution buffer: 1x PBS. Primary antibody: HA751998 (1/1,000, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature. Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature. |
|
Fig4:
Immunoprecipitation (IP) NUMB was immunoprecipitated in 0.2 mg U-87 MG (Human glioblastoma cell) cell lysate with HA751998 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751998 at 1/5,000 dilution. HRP Conjugated Rabbit IgG kappa light chain Mouse Monoclonal Antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: U-87 MG cell lysate (input) Lane 2: HA751998 IP in U-87 MG cell lysate Lane 3: Rabbit IgG instead of HA751998 in U-87 MG cell lysate Exposure time: 1 minute 18 seconds Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary dilution: HA751998, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature Predicted band size: 71 kDa Observed band size: 72/71/66/65 kDa |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.