ET1601-9

CD9 Recombinant Rabbit Monoclonal Antibody [SA35-08]

cat.: ET1601-9

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, IP, FC
Clonality:	Monoclonal
Clone number:	SA35-08

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: 25 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human CD9 aa 179-228 / 228.
Positive control:	HeLa cell lysate, K-562 cell lysate, MCF7 cell lysate, HCT 116 cell lysate, HepG2 cell lysate, SK-MEL-28 cell lysate, A375 cell lysate, B16-F1 cell lysate, SW480, CRC, human tonsil tissue, human spleen tissue, human kidney tissue, mouse brain tissue, mouse spleen tissue.
Subcellular location:	Cell membrane, Membrane
Recommended Dilutions: WB IF-Cell IHC-P IP FC	1:1,000-1:2,000 1:50 1:200-1:1,000 Use at an assay dependent concentration. 1:1,000
Uniprot #:	SwissProt: P21926 Human \| P40240 Mouse \| P40241 Rat
Alternative names:	Tetraspanin 29 5H9 5H9 antigen Antigen defined by monoclonal 602 29 Antigen defined by monoclonal 60229 BA-2/p24 antigen BA2 BTCC 1 BTCC1 CD9 CD9 antigen CD9 antigen p24 CD9 molecule CD9_HUMAN Cell growth inhibiting gene 2 protein Cell growth-inhibiting gene 2 protein DRAP 27 DRAP27 GIG2 Growth inhibiting gene 2 protein Leukocyte antigen MIC3 MIC3 Motility related protein Motility-related protein MRP 1 MRP-1 MRP1 p24 p24 antigen Tetraspanin-29 Tspan 29 Tspan-29 TSPAN29

Images

	Fig1: Western blot analysis of CD9 on different lysates with Rabbit anti-CD9 antibody (ET1601-9) at 1/2,000 dilution. Lane 1: HeLa cell lysate Lane 2: K-562 cell lysate Lane 3: MCF7 cell lysate Lane 4: HCT 116 cell lysate Lane 5: HepG2 cell lysate Lane 6: SK-MEL-28 cell lysate Lane 7: A375 cell lysate Lane 8: B16-F1 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 25 kDa Observed band size: 23 kDa Exposure time: 3 minutes 30 seconds; 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 (ET1601-9) 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.
	Fig2: ICC staining of CD9 in SW480 cells (green). 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 (ET1601-9, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig3: ICC staining of CD9 in CRC cells (green). 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 (ET1601-9, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

	Fig4: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CD9 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 (ET1601-9, 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-CD9 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 (ET1601-9, 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.
	Fig6: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-CD9 antibody (ET1601-9) 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 (ET1601-9) 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-CD9 antibody (ET1601-9) 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 (ET1601-9) 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.

Fig8: Flow cytometric analysis of B16-F1 cells labeling CD9.

Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (ET1601-9, 1μg/mL) (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”.