Human/Mouse/Rat Fascin Antibody

Catalog #: AF7745 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF7745
AF7745-SP

Detection of Human, Mouse, and Rat Fascin by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse/Rat Fascin Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects human, mouse, rat Fascin in direct ELISAs and Western blots. In direct ELISAs, less than 5% cross-reactivity with recombinant human Fascin-2 is observed.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human Fascin
Met1-Tyr493
Accession # Q16658

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

0.4 µg/mL

See below

Knockout Validated

Fascin is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in Fascin knockout HeLa 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, Mouse, and Rat Fascin by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, K562 human chronic myelogenous leukemia cell line, Neuro-2A mouse neuroblastoma cell line, and Rat-2 rat thymidine kinase-deficient embryonic fibroblast cell line. PVDF membrane was probed with 0.4 µg/mL of Sheep Anti-Human/Mouse/Rat Fascin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7745) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Fascin at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions using Immunoblot Buffer Group 1.

Knockout Validated

Western Blot Shows Human Fascin Antibody Specificity by Using Knockout Cell Line.

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Western Blot Shows Human Fascin Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Fascin knockout HeLa cell line (KO). PVDF membrane was probed with 0.4 µg/mL of Sheep Anti-Human/Mouse/Rat Fascin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7745) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Fascin at approximately 55 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1. New adjunct appears with knockout cell line.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

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: Fascin

Fascin (that which creates fascicles [bundles] of actin; also known as 55 kDa actin-bundling protein, p55 and singed-like protein) is an intracellular 55-58 kDa member of the fascin family of proteins. It has a restricted expression pattern, being found in oligodendrocytes, select endothelium, cerebellar stellate neurons and blood, interdigitating, and thymic medullary dendritic cells. Fascin is found associated with actin in filopodia, and serves to coordinate and stabilize actin bundle formation, both in normal cells and tumor cells. In the latter cell type, filopodia have been renamed invadopodia, and their appearance is crucial for the creation of a stable platform that coordinates local matrix degradation. Human fascin is 493 amino acids (aa) in length. It contains an N-terminal fascin-like domain (aa 139-256) that contains part of one of two actin-binding sequences (aa 136-143), followed by two additional fascin-like domains (aa 260-378 and 383-493), the latter of which contains the second actin-binding sequence (aa 386-395). There are also two acetylation sites and two utilized phosphorylation sites at Ser38 and Ser39. Phosphorylation of the latter site inhibits fascin interaction with actin. At least two isoform variants may exist. One contains a 12 aa substitution for aa 427-493, while a second shows a deletion of aa 371-426. Full-length human fascin shares 97% aa sequence identity with mouse fascin. Two additional human fascins termed retinal and testis fascin have been identified. They are products of distinct genes and share 56% and 27% aa sequence identity with the standard (p55) fascin, respectively.

Entrez Gene IDs

6624 (Human); 14086 (Mouse)

Alternate Names

FAN1; FAN1,55 kDa actin-bundling protein; fascin homolog 1, actin-bundling protein (Strongylocentrotus purpuratus); Fascin; FLJ38511; FSCN1; HSN; p55actin bundling protein; singed (Drosophila)-like (sea urchin fascin homolog like); singed-like (fascin homolog, sea urchin); Singed-like Protein; SNL; SNLfascin