Recombinant Anti-CD63 antibody [KILL150A] - BSA and Azide free (ab271296)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [KILL150A] to CD63 - BSA and Azide free
- Suitable for: WB, ICC/IF, Flow Cyt, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CD63 antibody [KILL150A] - BSA and Azide free
See all CD63 primary antibodies -
Description
Mouse monoclonal [KILL150A] to CD63 - BSA and Azide free -
Host species
Mouse -
Specificity
We recommend ab134045 for low expression samples.
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Tested applications
Suitable for: WB, ICC/IF, Flow Cyt, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Tissue, cells or virus. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: SK-MEL-28 and HUVEC whole cell lysate (boiled and un-boiled). Human lymph node tissue lysate. IHC-P: Human melanoma tissue. ICC/IF: SK-MEL-28 cells. Flow Cyt: SK-MEL-28
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General notes
ab271296 is the carrier-free version of ab271286.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
KILL150A -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab271296 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 26 kDa.
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ICC/IF |
Use at an assay dependent concentration.
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Flow Cyt |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 26 kDa. |
ICC/IF
Use at an assay dependent concentration. |
Flow Cyt
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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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All lanes : Anti-CD63 antibody [KILL150A] (ab271286) at 0.978 µg/ml
Lane 1 : SK-MEL-28 (human malignant melanoma), whole cell lysate
Lane 2 : HUVEC (human umbilical vein endothelial cell), whole cell lysate
Lane 3 : Human lymph node tissue lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Predicted band size: 26 kDaCD63 can undergo glycosylation as shown in lane 1, 2 and 3 (PMID: 9890706, 28740179).
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure times: Lanes 1-2: 3 mins; Lane 3: 48 secs.
This blot was developed using a higher sensitivity ECL substrate.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab271286).
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All lanes : Anti-CD63 antibody [KILL150A] (ab271286) at 1/1000 dilution
Lanes 1 & 5 : Un-boiled HUVEC (human umbilical vein endothelial cell) whole cell lysate
Lanes 2 & 6 : Un-boiled SK-MEL-28 (human malignant melanoma) whole cell lysate
Lane 3 : Boiled HUVEC (human umbilical vein endothelial cell) whole cell lysate
Lane 4 : Boiled SK-MEL-28 (human malignant melanoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution
Developed using the ECL technique.
Predicted band size: 26 kDa
Observed band size: 30-60 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM /TBST
Exposure time: Lane1-4: 180s; Lane 5-6: 80s
We recommend not boiling lysate before loading onto the gel and using a higher sensitive ECL substrate to increase the band intensity.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab271286).
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Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling CD63 with ab271286 at 1/4000 dilution (0.978µg/ml) followed by ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879). Cytoplasmic staining on human melanoma. The section was incubated with ab271268 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab271286).
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Immunohistochemical analysis of paraffin-embedded human breast tissue labeling CD63 with ab271286 at 1/4000 dilution (0.978µg/ml) followed by ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879). The section was incubated with ab271268 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Negative control: Nearly no staining on human breast.
Secondary antibody only control: Secondary antibody is ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab271286).
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SK-MEL-28 cells labelling CD63 with ab271286 at 1/50 dilution (19.56µg/ml), followed by ab150113 Goat Anti-mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing lysosome/endosomes (ab252919) co-staining in SK-MEL-28 cells.
ab252919 Anti-CD63 rabbit monoclonal antibody was used to counterstain tubulin at 1/250 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at a 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).
Negative control 1: ab271286 at a 1/50 dilution (19.56µg/ml) followed by ab150080 at a 1/1000 dilution.
Negative control 2: ab259919 at a 1/200 dilution followed by ab150113 at a 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab271286).
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Flow cytometric analysis of SK-MEL-28 (human malignant melanoma) cells labelling CD63 with ab271286 at 1/1000 dilution (19.56µg/ml) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab271286).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
References (0)
ab271296 has not yet been referenced specifically in any publications.