CD63 Rabbit mAb - SAB | Signalway Antibody

Catalog No.	Package Size	Price	Quantity
62246-1	100ul	$599.00

Datasheet Images Protocol

Description

Product NameCD63 Rabbit mAb
Host SpeciesRabbit
ClonalityMonoclonal
Clone No.SR4552
PurificationProtein A affinity purified.
ApplicationsWB;IHC;IF;IP;FC
Species ReactivityHuman
Immunogen TypeProtein
Immunogen DescriptionRecombinant protein within
Conjugatesunconjugated
Target NameCD63
UniprotP08962
Calculated MW 26 kDa
Concentration 1ug/ul
Formulation PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Storage Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Application Details

Images

Western blot analysis of CD63 on different lysates with Rabbit CD63 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 ug/Lane.
Predicted band size: 26 kDa
Observed band size: 30-65 kDa
Immunohistochemical analysis of paraffin-embedded human melanoma tissue with CD63 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.
Immunocytochemistry analysis of K-562 cells labeling CD63 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 CD63 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.
Flow cytometric analysis of K-562 cells labeling CD63.
Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (1ug/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).
CD63 was immunoprecipitated from 0.2 mg SK-MEL-28 cell lysate at 2 ug/25 ul agarose. Western blot was performed from the immunoprecipitate 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: CD63 IP in SK-MEL-28 cell lysate
Lane 3: Rabbit IgG instead of CD63 in SK-MEL-28 cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST

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CD63 Rabbit mAb #62246

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