Human Numb Antibody

Catalog #: AF4338 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF4338
AF4338-SP

Detection of Human Numb by Western Blot.

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Human Numb Antibody Summary

Species Reactivity

Human

Specificity

Detects human Numb in direct ELISAs and Western blots.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human Numb
Thr356-Leu592
Accession # AAH68476

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunocytochemistry

5-15 µg/mL

Immersion fixed RT‑4 human urinary bladder transitional cell papilloma cell line

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human Numb by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line, A549 human lung carcinoma cell line, and HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Numb Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4338) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Numb at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

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Numb in RT‑4 Human Cell Line. Numb was detected in immersion fixed RT‑4 human urinary bladder transitional cell papilloma cell line using Sheep Anti-Human Numb Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4338) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Numb

Numb is a member of a family of cytoplasmic adaptor proteins termed CLASPS (Clathran-associated Sorting Proteins). These link transmembrane cargo proteins to the endocytic machinery. Numb, a conserved cell-fate determining factor, is normally associated with the basolateral surface of polarized cells. Movement towards the apical surface induces its phosphorylation, which precludes its entry into this region. Human Numb is 651 amino acids (aa) in length. It contains one PTB (aa 24‑172) and one PRR (aa 366‑413) domain. There is a potential for multiple alternate splice forms that involve all regions of the molecule. Over aa 356‑592, human Numb shares 88% and 90% aa identity with mouse and canine Numb, respectively.

Entrez Gene IDs

8650 (Human); 18222 (Mouse); 29419 (Rat)

Alternate Names

h-Numb; mnb; m-numb; numb (Drosophila) homolog; numb homolog (Drosophila); Numb; protein numb homolog; Protein S171; S171