Anti-CD63 antibody (ab68418)
Key features and details
- Rabbit polyclonal to CD63
- Suitable for: WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-CD63 antibody
See all CD63 primary antibodies -
Description
Rabbit polyclonal to CD63 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species WB Human
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Immunogen
Synthetic peptide corresponding to Human CD63 aa 100-200 conjugated to keyhole limpet haemocyanin.
(Peptide available asab93074) -
Positive control
- This antibody gave a positive signal in Human platelet and HepG2 whole cell lysates.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab68418 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species WB HumanApplication Abreviews Notes WB (1) Use a concentration of 1 µg/ml. Detects a band of approximately 24,43 kDa (predicted molecular weight: 25 kDa).Notes WB
Use a concentration of 1 µg/ml. Detects a band of approximately 24,43 kDa (predicted molecular weight: 25 kDa).Target
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Function
This antigen is associated with early stages of melanoma tumor progression. May play a role in growth regulation. -
Tissue specificity
Dysplastic nevi, radial growth phase primary melanomas, hematopoietic cells, tissue macrophages. -
Sequence similarities
Belongs to the tetraspanin (TM4SF) family. -
Cellular localization
Cell membrane. Lysosome membrane. Late endosome membrane. Also found in Weibel-Palade bodies of endothelial cells. Located in platelet dense granules. - Information by UniProt
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Database links
- Entrez Gene: 967 Human
- Omim: 155740 Human
- SwissProt: P08962 Human
- Unigene: 445570 Human
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Alternative names
- Lysosomal associated membrane protein 3 antibody
- CD 63 antibody
- CD63 antibody
see all
Images
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Western blot - Anti-CD63 antibody (ab68418)All lanes : Anti-CD63 antibody (ab68418) at 1 µg/ml
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Wild-type HAP1 Breferldin A treated whole cell lysate
Lane 3 : CD63 knockout HAP1 whole cell lysate
Lane 4 : CD63 knockout HAP1 Breferldin A treated whole cell lysate
Lane 5 : HL60 whole cell lysate
Lane 6 : Human platelets whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 25 kDaLanes 1 - 6: Merged signal (red and green). Green - ab68418 observed at 26 kDa. Red - loading control, ab8245, observed at 38 kDa.
ab68418 was shown to recognize CD63 in wild-type HAP1 as signal was lost at the expected MW in CD63 knockout HAP1 cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CD63 knockout samples were subjected to SDS-PAGE. Ab68418 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 µg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-CD63 antibody (ab68418)Anti-CD63 antibody (ab68418) at 1 µg/ml + Platelet (Human) Whole Cell Lysate at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 43 kDa why is the actual band size different from the predicted?
Exposure time: 8 minutesThis blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab68418 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
CD63 has a predicted molecular weight of 25 kDa; however it has a number of potential glycosylation sites which may affect the migration of the protein (SwissProt data). This is consistent with the literature which describes a band between 30-60 kDa in human platelets(PMID:1993697).
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Western blot - Anti-CD63 antibody (ab68418)Anti-CD63 antibody (ab68418) at 1 µg/ml + HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?
Additional bands at: 145 kDa, 42 kDa (possible glycosylated form). We are unsure as to the identity of these extra bands.
Exposure time: 20 minutesThe 42 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to human CD63 antigen.
Datasheets and documents
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SDS download
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Datasheet download
References (49)
ab68418 has been referenced in 49 publications.
- Hu H et al. Exosome-Derived miR-486-5p Regulates Cell Cycle, Proliferation and Metastasis in Lung Adenocarcinoma via Targeting NEK2. Front Bioeng Biotechnol 8:259 (2020). PubMed: 32322578
- Chen F et al. Hypoxia induced changes in miRNAs and their target mRNAs in extracellular vesicles of esophageal squamous cancer cells. Thorac Cancer 11:570-580 (2020). PubMed: 31922357
- Moloney BM et al. Investigating the Potential and Pitfalls of EV-Encapsulated MicroRNAs as Circulating Biomarkers of Breast Cancer. Cells 9:N/A (2020). PubMed: 31936142
- Uysal-Onganer P et al. Peptidylarginine Deiminase Isozyme-Specific PAD2, PAD3 and PAD4 Inhibitors Differentially Modulate Extracellular Vesicle Signatures and Cell Invasion in Two Glioblastoma Multiforme Cell Lines. Int J Mol Sci 21:N/A (2020). PubMed: 32098295
- Sancandi M et al. Protein Deimination Signatures in Plasma and Plasma-EVs and Protein Deimination in the Brain Vasculature in a Rat Model of Pre-Motor Parkinson's Disease. Int J Mol Sci 21:N/A (2020). PubMed: 32326590
Images
-
Western blot - Anti-CD63 antibody (ab68418)All lanes : Anti-CD63 antibody (ab68418) at 1 µg/ml
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Wild-type HAP1 Breferldin A treated whole cell lysate
Lane 3 : CD63 knockout HAP1 whole cell lysate
Lane 4 : CD63 knockout HAP1 Breferldin A treated whole cell lysate
Lane 5 : HL60 whole cell lysate
Lane 6 : Human platelets whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 25 kDaLanes 1 - 6: Merged signal (red and green). Green - ab68418 observed at 26 kDa. Red - loading control, ab8245, observed at 38 kDa.
ab68418 was shown to recognize CD63 in wild-type HAP1 as signal was lost at the expected MW in CD63 knockout HAP1 cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CD63 knockout samples were subjected to SDS-PAGE. Ab68418 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 µg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
-
Western blot - Anti-CD63 antibody (ab68418)Anti-CD63 antibody (ab68418) at 1 µg/ml + Platelet (Human) Whole Cell Lysate at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 43 kDa why is the actual band size different from the predicted?
Exposure time: 8 minutesThis blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab68418 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
CD63 has a predicted molecular weight of 25 kDa; however it has a number of potential glycosylation sites which may affect the migration of the protein (SwissProt data). This is consistent with the literature which describes a band between 30-60 kDa in human platelets(PMID:1993697).
-
Western blot - Anti-CD63 antibody (ab68418)Anti-CD63 antibody (ab68418) at 1 µg/ml + HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?
Additional bands at: 145 kDa, 42 kDa (possible glycosylated form). We are unsure as to the identity of these extra bands.
Exposure time: 20 minutesThe 42 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to human CD63 antigen.
