CD63 Recombinant Rabbit Monoclonal Antibody [PSH06-64]
cat.: HA722731
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, FC, IP, IHC-P |
| Clonality: | Monoclonal |
| Clone number: | PSH06-64 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 26 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within |
| Positive control: | SK-MEL-28 cell lysate, U-87 MG cell lysate, HUVEC cell lysate, K-562 cell lysate, HEK-293 cell lysate, HL-60 cell lysate, K-562, human melanoma tissue. |
| Subcellular location: | Cell membrane, Lysosome membrane, Late endosome membrane, Endosome, multivesicular body, Melanosome, Secreted, extracellular exosome, Cell surface. |
| Recommended Dilutions:
WB IF-Cell FC IP IHC-P |
1:1,000 1:250 1:1,000 1-2μg/sample 1:200 |
| Uniprot #: | SwissProt: P08962 Human |
| Alternative names: | Lysosomal associated membrane protein 3 CD 63 CD63 CD63 antigen (melanoma 1 antigen) CD63 antigen CD63 antigen melanoma 1 antigen CD63 molecule CD63_HUMAN gp55 Granulophysin LAMP 3 LAMP-3 LAMP3 LIMP Lysosomal-associated membrane protein 3 Lysosome associated membrane glycoprotein 3 Lysosome-associated membrane glycoprotein 3 Mast cell antigen AD1 ME491 Melanoma 1 antigen Melanoma associated antigen ME491 Melanoma associated antigen MLA1 Melanoma-associated antigen ME491 MGC72893 MLA 1 MLA1 NGA Ocular melanoma associated antigen Ocular melanoma-associated antigen OMA81H PTLGP40 Tetraspanin 30 Tetraspanin-30 Tspan 30 Tspan-30 TSPAN30 |
Images
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Fig1:
Western blot analysis of CD63 on different lysates with Rabbit anti-CD63 antibody (HA722731) at 1/1,000 dilution. Lane 1: SK-MEL-28 cell lysate Lane 2: Jurkat cell lysate (negative) Lane 3: U-87 MG cell lysate Lane 4: HUVEC cell lysate Lane 5: K-562 cell lysate Lane 6: HEK-293 cell lysate Lane 7: HL-60 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 26 kDa Observed band size: 30-65 kDa Exposure time: Lane 1: 4 seconds; Lane 2-7: 18 seconds; ECL: K1802; 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 (HA722731) 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. |
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Fig2:
Immunocytochemistry analysis of K-562 cells labeling CD63 with Rabbit anti-CD63 antibody (HA722731) at 1/250 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-CD63 antibody (HA722731) at 1/250 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. |
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Fig3:
Immunohistochemical analysis of paraffin-embedded human melanoma tissue with Rabbit anti-CD63 antibody (HA722731) 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 (HA722731) 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. |
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Fig4:
Flow cytometric analysis of K-562 cells labeling CD63. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA722731, 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). |
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Fig5:
CD63 was immunoprecipitated from 0.2 mg SK-MEL-28 cell lysate with HA722731 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722731 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: SK-MEL-28 cell lysate (input) Lane 2: HA722731 IP in SK-MEL-28 cell lysate Lane 3: Rabbit IgG instead of HA722731 in SK-MEL-28 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 1 minute 31 seconds; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.