ET1705-18

Fascin Recombinant Rabbit Monoclonal Antibody [JM12-53]

cat.: ET1705-18

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JM12-53

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 54 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human Fascin aa 221-389 / 493.
Positive control:	HeLa cell lysate, Mouse brain tissue lysate, Mouse kidney tissue lysate, Mouse heart tissue lysate, Rat brain tissue lysate, SH-SY5Y, human kidney tissue, rat brain tissue, human tonsil tissue.
Subcellular location:	Cytoskeleton, stress fiber, cytosol, cell cortex, filopodium, invadopodium, microvillus, cell junction.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC	1:1,000-1:2,000 1:2,000 1:50-1:200 1:1,000 1:1,000
Uniprot #:	SwissProt: Q16658 Human \| Q61553 Mouse \| P85845 Rat
Alternative names:	55 kDa actin bundling protein 55 kDa actin-bundling protein Actin bundling protein actin bundling protein, 55-KD FAN 1 FAN1 Fascin 1 Fascin actin bundling protein 1 Fascin Fascin homolog 1 actin bundling protein (Strongylocentrotus purpuratus) Fascin homolog 1 Fascin, sea urchin, homolog of, 1 Fascin1 FLJ38511 FSCN 1 FSCN1 FSCN1_HUMAN HSN p55 Singed (Drosophila) like (sea urchin fascin homolog like) Singed drosophila homolog like Singed like (fascin homolog sea urchin) (Drosophila) Singed like (fascin homolog sea urchin) Singed like protein Singed, drosophila, homolog of Singed-like protein SNL Strongylocentrotus purpuratus

Images

Fig1: Western blot analysis of Fascin on different lysates with Rabbit anti-Fascin antibody (ET1705-18) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: Mouse brain tissue lysate
Lane 3: Mouse kidney tissue lysate
Lane 4: Mouse heart tissue lysate
Lane 5: Rat brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 54 kDa
Observed band size: 54 kDa

Exposure time: 4 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-18) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of Fascin on different lysates with Rabbit anti-Fascin antibody (ET1705-18) at 1/1,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-Fascin KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 54 kDa
Observed band size: 54 kDa

Exposure time: 4 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-18) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of SH-SY5Y cells labeling Fascin with Rabbit anti-Fascin antibody (ET1705-18) at 1/2,000 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Fascin antibody (ET1705-18) at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig4: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Fascin antibody (ET1705-18) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1705-18) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig5: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Fascin antibody (ET1705-18) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1705-18) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig6: Flow cytometric analysis of SH-SY5Y cells labeling Fascin. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1705-18, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.