ET7108-95

Nectin 2 Recombinant Rabbit Monoclonal Antibody [JG39-64]

cat.: ET7108-95

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JG39-64

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. 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: 58 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human Nectin 2 aa 370-538 / 538.
Positive control:	HT-29 cell lysate, A549 cell lysate, MDA-MB-231 cell lysate, NIH/3T3 cell lysate, Mouse testis tissue lysate, Rat testis tissue lysate, MCF7, human testis tissue, mouse testis tissue, rat testis tissue.
Subcellular location:	Cell membrane.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:5,000-1:10,000 1:100 1:200 1:1:1,000
Uniprot #:	SwissProt: Q92692 Human \| P32507 Mouse \| Q5FVC5 Rat
Alternative names:	CD 112 CD112 CD112 antigen Herpes virus entry mediator B Herpes virus entry protein B Herpesvirus entry mediator B Herpesvirus entry protein B Hve B HveB Nectin-2 Nectin2 Poliovirus receptor like 2 Poliovirus receptor related 2 (herpesvirus entry mediator B) Poliovirus receptor related 2 Poliovirus receptor related protein 2 Poliovirus receptor-related protein 2 PRR 2 PRR2 PVRL 2 PVRL2 PVRL2_HUMAN PVRR 2 PVRR2

Images

Fig1: Western blot analysis of Nectin 2 on different lysates with Rabbit anti-Nectin 2 antibody (ET7108-95) at 1/5,000 dilution.

Lane 1: HT-29 cell lysate
Lane 2: A549 cell lysate
Lane 3: MDA-MB-231 cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: Mouse testis tissue lysate
Lane 6: Rat testis tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 58 kDa
Observed band size: 75 kDa

Exposure time: 16 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 (ET7108-95) at 1/5,000 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 Nectin 2 on different lysates with Rabbit anti-Nectin 2 antibody (ET7108-95) at 1/10,000 dilution.

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

Lysates/proteins at 10 µg/Lane.

Predicted band size: 58 kDa
Observed band size: 70-85 kDa

Exposure time: 10 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 (ET7108-95) at 1/10,000 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 MCF7 cells labeling Nectin 2 with Rabbit anti-Nectin 2 antibody (ET7108-95) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Nectin 2 antibody (ET7108-95) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-Nectin 2 antibody (ET7108-95) 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 (ET7108-95) 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 mouse testis tissue with Rabbit anti-Nectin 2 antibody (ET7108-95) 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 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7108-95) at 1/200 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: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Nectin 2 antibody (ET7108-95) 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 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7108-95) at 1/200 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.

Fig7: Flow cytometric analysis of MCF7 cells labeling Nectin 2.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET7108-95, 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”.