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| Product Name | CD63 Recombinant Rabbit Monoclonal Antibody |
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| Antibody Type | Primary Antibodies |
| Product description | The tetraspanins are integral membrane proteins expressed on cell surface and granular membranes of hematopoietic cells and are components of multi-molecular complexes with specific integrins. The tetraspanin CD63 (also known as LAMP-3, Melanoma-associated antigen ME491, TSPAN30, MLA1 and OMA81H) is a lysosomal membrane glycoprotein that translocates to the plasma membrane after platelet activation. CD63 is expressed on activated platelets, monocytes and macrophages, and is weakly expressed on granulocytes, T cell and B cells. It is located on the basophilic granule membranes and on the plasma membranes of lymphocytes and granulocytes. CD63 is a member of the TM4 superfamily of leukocyte glycoproteins that includes CD9, CD37 and CD53, which contain four transmembrane regions. CD63 may play a role in phagocytic and intracellular lysosome-phagosome fusion events. CD63 deficiency is associated with Hermansky-Pudlak syndrome. |
| Immunogen | Recombinant protein within |
| Clonality | monoclonal |
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| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:1000 IHC:1:200 ICC/IF:1:250 FC:1:1000 IP:1-2μg/sample | |
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | Lysosomal associated membrane protein 3 antibody CD 63 antibody CD63 antibody CD63 antigen (melanoma 1 antigen) antibody CD63 antigen antibody CD63 antigen melanoma 1 antigen antibody CD63 molecule antibody CD63_HUMAN antibody gp55 antibody Granulophysin antibody LAMP 3 antibody LAMP-3 antibody LAMP3 antibody LIMP antibody Lysosomal-associated membrane protein 3 antibody Lysosome associated membrane glycoprotein 3 antibody Mast cell antigen AD1 antibody ME491 antibody Melanoma 1 antigen antibody Melanoma associated antigen ME491 antibody Melanoma associated antigen MLA1 antibody Melanoma-associated antigen ME491 antibody MGC72893 antibody MLA 1 antibody MLA1 antibody NGA antibody Ocular melanoma associated antigen antibody Ocular melanoma-associated antigen antibody OMA81H antibody PTLGP40 antibody Tetraspanin 30 antibody Tetraspanin-30 antibody Tspan 30 antibody Tspan-30 antibody TSPAN30 antibody |
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| Molecular Weight(MW) | 26 kDa(Observed band size: 30-65 kDa) |
| Cellular Localization | Cell membrane, Lysosome membrane, Endosome, Melanosome, Secreted. |
| SwissProt ID | P08962 |
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WB
Western blot analysis of CD63 on different lysates with Rabbit anti-CD63 antibody 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. Exposure time: Lane 1: 4 seconds; Lane 2-7: 18 seconds; Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human melanoma tissue with Rabbit anti-CD63 antibod 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 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.
ICC/IF
Immunocytochemistry analysis of K-562 cells labeling CD63 with Rabbit anti-CD63 antibody 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 at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of K-562 cells labeling CD63. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
CD63 was immunoprecipitated from 0.2 mg SK-MEL-28 cell lysate with Rabbit anti-CD63 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-CD63 antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: SK-MEL-28 cell lysate (input), Lane 2: Rabbit anti-CD63 antibody IP in SK-MEL-28 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-CD63 antibody in SK-MEL-28 cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 1 minute 31 seconds.| Application Notes | WB:1:1000 IHC:1:200 ICC/IF:1:250 FC:1:1000 IP:1-2μg/sample |
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| Form | Liquid |
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| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
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