ET1702-91

ARF6 Recombinant Rabbit Monoclonal Antibody [JF101-2]

cat.: ET1702-91

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, IHC-Fr
Clonality:	Monoclonal
Clone number:	JF101-2

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: 20 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human ARF6 aa 133-165 / 175.
Positive control:	Human liver tissue lysates, Hela, MCF-7, HepG2, human stomach tissue, mouse embryonic brain tissue.
Subcellular location:	Golgi apparatus, Cell membrane, Endosome membrane, Cell projection, Midbody, Cytoplasm, Cleavage furrow.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P IHC-Fr	1:500-1:2,000 1:100-1:500 1:200 1:50-1:200 1:500
Uniprot #:	SwissProt: P62330 Human \| P62331 Mouse \| P62332 Rat
Alternative names:	ADP ribosylation factor 6 ADP ribosylation factor protein 6 ADP-ribosylation factor 6 ARF6 ARF6_HUMAN DKFZp564M0264 Small GTP binding protein Small GTPase

Images

	Fig1: Immunofluorescence analysis of frozen mouse embryo tissue with Rabbit anti-ARF6 antibody (ET1702-91) at 1/500 dilution. The section was not undergone antigen retrieval. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1702-91, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig2: Immunofluorescence analysis of frozen mouse embryonic brain tissue with Rabbit anti-ARF6 antibody (ET1702-91) at 1/500 dilution. The section was not undergone antigen retrieval. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1702-91, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig3: Western blot analysis of ARF6 on different lysates with Rabbit anti-ARF6 antibody (ET1702-91) at 1/1,000 dilution. Lane 1: A549-WT cell lysate Lane 2: A549-KD ARF6 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 20 kDa Observed band size: 18 kDa Exposure time: 3 minutes; 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 (ET1702-91) 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.

	Fig4: Western blot analysis of ARF6 on human liver tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1702-91, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.
	Fig5: ICC staining of ARF6 in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-91, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig6: ICC staining of ARF6 in MCF-7 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-91, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig7: ICC staining of ARF6 in HepG2 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-91, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Fig8: Immunohistochemical analysis of paraffin-embedded human stomach tissue using anti-ARF6 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1702-91, 1/200) for 30 minutes 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.

Fig9: Immunofluorescence analysis of paraffin-embedded mouse embryonic brain tissue labeling ARF6 with Rabbit anti-ARF6 antibody (ET1702-91) at 1/200 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1702-91, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

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