ET1610-33

Islet 1 Recombinant Rabbit Monoclonal Antibody [SC05-64]

cat.: ET1610-33

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC, IP, IHC-Fr
Clonality:	Monoclonal
Clone number:	SC05-64

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: 39 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human Islet 1 aa 262-205 / 349.
Positive control:	SH-SY5Y cell lysate, Neuro-2a cell lysate, PC-12 cell lysate, SH-SY5Y, Neuro-2a, human pancreas tissue, mouse embryo tissue, rat pancreas tissue.
Subcellular location:	Nucleus.
Recommended Dilutions: WB IF-Cell IHC-P FC IP IHC-Fr	1:2,000 1:100 1:1,000 1:1,000 1-2μg/sample 1:500
Uniprot #:	SwissProt: P61371 Human \| P61372 Mouse \| P61374 Rat
Alternative names:	Insulin gene enhancer protein ISL 1 Insulin gene enhancer protein ISL-1 Insulin related protein ISL 1 ISL LIM homeobox 1 ISL1 ISL1 transcription factor LIM homeodomain ISL1 transcription factor, LIM/homeodomain (islet 1) ISL1 transcription factor, LIM/homeodomain ISL1_HUMAN Islet-1 Islet1

Images

	Fig1: Immunofluorescence analysis of frozen mouse embryo tissue with Rabbit anti-Islet 1 antibody (ET1610-33) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. 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 (ET1610-33, 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-Islet 1 antibody (ET1610-33) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. 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 (ET1610-33, 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: Immunofluorescence analysis of frozen mouse embryonic brain tissue with Rabbit anti-Islet 1 antibody (ET1610-33) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. 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 (ET1610-33, 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).

Fig4: Immunofluorescence analysis of frozen mouse pancreas tissue with Rabbit anti-Islet 1 antibody (ET1610-33) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. 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 (ET1610-33, 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).

Fig5: Western blot analysis of Islet 1 on different lysates with Rabbit anti-Islet 1 antibody (ET1610-33) at 1/2,000 dilution.

Lane 1: SH-SY5Y cell lysate (5 µg/Lane)
Lane 2: Neuro-2a cell lysate (30 µg/Lane)
Lane 3: PC-12 cell lysate (30 µg/Lane)

Predicted band size: 39 kDa
Observed band size: 45 kDa

Exposure time: Lane 1: 10 seconds; Lane 2-3: 1 minute; 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 (ET1610-33) at 1/2,000 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.

Fig6: Immunocytochemistry analysis of SH-SY5Y cells labeling Islet 1 with Rabbit anti-Islet 1 antibody (ET1610-33) at 1/100 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-Islet 1 antibody (ET1610-33) at 1/100 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 (M1305-2, 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.

Fig7: Immunocytochemistry analysis of Neuro-2a cells labeling Islet 1 with Rabbit anti-Islet 1 antibody (ET1610-33) at 1/100 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-Islet 1 antibody (ET1610-33) at 1/100 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 (M1305-2, 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.

	Fig8: Immunohistochemical analysis of paraffin-embedded human pancreas tissue with Rabbit anti-Islet 1 antibody (ET1610-33) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1610-33) 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.
	Fig9: Immunohistochemical analysis of paraffin-embedded mouse embryo tissue with Rabbit anti-Islet 1 antibody (ET1610-33) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1610-33) 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.
	Fig10: Immunohistochemical analysis of paraffin-embedded rat pancreas tissue with Rabbit anti-Islet 1 antibody (ET1610-33) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (ET1610-33) 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.

Fig11: Flow cytometric analysis of SH-SY5Y cells labeling Islet 1.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1610-33, 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”.