Invitrogen

NUMB Polyclonal Antibody

Product Image Gallery

NUMB Antibody in Immunocytochemistry (ICC/IF) — 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 # PA5-21143) at 5µg/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.

NUMB Antibody in Immunohistochemistry (IHC) — Immunohistochemistry of NUMB in human lung tissue with NUMB Polyclonal Antibody (Product # PA5-21143) at 5 µg/mL.

NUMB Antibody in Western Blot (WB) — Western blot was performed using Anti-NUMB Polyclonal Antibody (Product # PA5-21143) on whole cell extracts (30 µg lysate) of U-87 MG (Lane1), PC-3 (Lane 2), DU 145 (Lane 3), A549 (Lane 4), SH-SY5Y(Lane 5), HeLa (Lane 6), Mouse Lungs (Lane 7), Mouse Colon (Lane 8), Mouse Heart (Lane 9) and Mouse Kidney (Lane 10)and NUMB major isoform 65 kDa was observed along with other isoforms between 53-70 kDa in all samples except Mouse Colon, Heart and Kidney. 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 Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Product Details

PA5-21143

Applications	Tested Dilution	Publications
Western Blot (WB)	0.25-0.5 µg/mL	-
Immunohistochemistry (IHC)	5 µg/mL	-
Immunocytochemistry (ICC/IF)	5-20 µg/mL	-

Product Specifications
Species Reactivity	Human, Mouse, Rat
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	A 17 amino acid peptide near the carboxy terminus of human NUMB. View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS
Contains	0.02% sodium azide
Storage conditions	Maintain refrigerated at 2-8°C for up to 3 months. For long term storage store at -20°C
RRID	AB_11154464

Product Specific Information

A suggested positive control is mouse lung tissue lysate.

PA5-21143 can be used with blocking peptide PEP-1257.

Target Information

Protein NUMB homolog is an evolutionarily conserved protein that controls the differentiation of neuronal progenitor cells. Recent studies have revealed that NUMB functions in clathrin-dependent endocytosis by binding to the AP-2 complex. NUMB is also expressed in postmitotic neurons and plays a role in axonal growth. Neural cells expressing Numb isoforms with short phosphotyrosine-binding (SPTB) domain undergo extensive neurite outgrowth, an effect that can be blocked by voltage-gated Ca2+ channel (VGCC) inhibitor or by Ca2+ chelator.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.