NUMB Recombinant Rabbit Monoclonal Antibody [PSH25-86] - BSA and Azide free
Catalog# HA751998
NUMB Recombinant Rabbit Monoclonal Antibody [PSH25-86] - BSA and Azide free
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WB
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FC(Intra)
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IP
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Human
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Rat
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Green monkey
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unconjugated
Overview
Product Name
NUMB Recombinant Rabbit Monoclonal Antibody [PSH25-86] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human NUMB aa 301-650.
Species Reactivity
Human, Rat, Green monkey
Validated Applications
WB, FC(Intra), IP
Target Molecular Weight
Predicted band size: 71 kDa
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.
Conjugation
unconjugated
Clone Number
PSH25-86
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:10,000
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FC(Intra)
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1:1,000
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IP
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1-2μg/sample
Target
Function
Protein numb homolog is a protein that in humans is encoded by the NUMB gene. The protein encoded by this gene plays a role in the determination of cell fates during development. The encoded protein, whose degradation is induced in a proteasome-dependent manner by MDM2, is a membrane-bound protein that has been shown to associate with EPS15, LNX1, and NOTCH1. Four transcript variants encoding different isoforms have been found for this gene. The protein Numb is coded for by the gene, NUMB, whose mechanism appears to be evolutionarily conserved. Numb has been extensively studied in both invertebrates and mammals, though its function is best understood in Drosophila. Numb plays a crucial role in asymmetrical cell division during development, allowing for differential cell fate specification in the central and peripheral nervous systems. During neurogenesis, Numb localizes to one side of the mother cell such that it is distributed selectively to one daughter cell. This asymmetric division allows a daughter cell containing Numb to acquire a different fate than the other daughter cell.
Background References
1. Kudinov AE et al. Musashi-2 (MSI2) supports TGF- signaling and inhibits claudins to promote non-small cell lung cancer (NSCLC) metastasis. Proc Natl Acad Sci U S A 113:6955-60 (2016).
2. Zhao C et al. Numb family proteins are essential for cardiac morphogenesis and progenitor differentiation. Development 141:281-95 (2014).
Subcellular Location
Cell membrane, Endosome membrane.
Synonyms
c14_5527 antibody
C14orf41 antibody
FLJ31314 antibody
h Numb antibody
h-Numb antibody
Numb antibody
Numb homolog (Drosophila) antibody
Numb homolog antibody
Numb protein homolog antibody
NUMB_HUMAN antibody
ExpandImages
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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 -
☑ Relative expression (RE)
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 -
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. -
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
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"