Recombinant Anti-Ly-6K antibody [EPR23021-61] (ab246486)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ly-6K antibody [EPR23021-61] (ab246486)](https://www.abcam.com/ps/products/246/ab246486/Images/ab246486-1-anti-ly6k-antibody-epr2302161-immunohistochemistry-human-testis.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ly-6K antibody [EPR23021-61] (ab246486)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23021-61] to Ly-6K
- Suitable for: WB, IHC-P, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Ly-6K antibody [EPR23021-61]
See all Ly-6K primary antibodies -
Description
Rabbit monoclonal [EPR23021-61] to Ly-6K -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IPmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa cell lysate; Human testis tissue lysate. IHC-P:Human testis tissue. IP: Human testis tissue lysate.
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General notes
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23021-61 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab246486 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 |
|---|---|---|
| WB |
1/1000. Detects a band of approximately 24 kDa (predicted molecular weight: 19 kDa).
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| IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| IP |
1/30.
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| Notes |
|---|
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WB
1/1000. Detects a band of approximately 24 kDa (predicted molecular weight: 19 kDa). |
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IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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IP
1/30. |
Target
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Relevance
The Lymphocyte Antigen 6 Complex is a 15 kDa phosphatidyl inositol anchored molecule, attached to the cell membrane by a GPI-anchor. The Lymphocyte Antigen 6 Complex is present on most immature T cells of the thymus, on lymphoid cells in bone marrow and peripheral B cells, but absent on peripheral T cells. -
Cellular localization
Cell Membrane -
Database links
- Entrez Gene: 54742 Human
- Omim: 615093 Human
- SwissProt: Q17RY6 Human
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Alternative names
- Lymphocyte antigen Ly-6K antibody
- 2410015A16Rik antibody
- 3110035B01Rik antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ly-6K antibody [EPR23021-61] (ab246486)
Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Ly-6K with ab246486 at 1/2000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in spermatocytes of human testis (PMID:18089789). The section was incubated with ab246486 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 a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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![Western blot - Anti-Ly-6K antibody [EPR23021-61] (ab246486)](https://www.abcam.com/ps/products/246/ab246486/Images/ab246486-448053-anti-ly-6k-antibody-epr23021-61-western-blot-human-knockout-lysate.jpg)
Western blot - Anti-Ly-6K antibody [EPR23021-61] (ab246486)All lanes : Anti-Ly-6K antibody [EPR23021-61] (ab246486) at 1/1000 dilution
Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : LY6K knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 19 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?Lanes 1-2: Merged signal (red and green). Green - ab246486 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa.
ab246486 Anti-Ly-6K antibody [EPR23021-61] was shown to specifically react with Ly-6K in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265874 (knockout cell lysate ab263248) was used. Wild-type and Ly-6K knockout samples were subjected to SDS-PAGE. ab246486 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-Ly-6K antibody [EPR23021-61] (ab246486)](https://www.abcam.com/ps/products/246/ab246486/Images/ab246486-388952-anti-ly6k-antibody-epr2302161-western-blot-human-testis-human-heart-lysates.jpg)
Western blot - Anti-Ly-6K antibody [EPR23021-61] (ab246486)All lanes : Anti-Ly-6K antibody [EPR23021-61] (ab246486) at 1/1000 dilution
Lane 1 : Human testis tissue lysate
Lane 2 : Human heart tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution
Predicted band size: 19 kDa
Observed band size: 24 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile observed is consistent with what has been described in the literature (PMID: 27197181, 18089789).
Negative control: Human heart (PMID:18089789) Exposure time: 10 seconds
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![Immunoprecipitation - Anti-Ly-6K antibody [EPR23021-61] (ab246486)](https://www.abcam.com/ps/products/246/ab246486/Images/ab246486-3-anti-ly6k-antibody-epr2302161-immunoprecipitation-human-testis-tissue-lysate-10-ug.jpg)
Immunoprecipitation - Anti-Ly-6K antibody [EPR23021-61] (ab246486)Ly-6K was immunoprecipitated from 0.35 mg Human testis tissue lysate with ab246486 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab246486 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/1000 dilution.
Lane 1: Human testis tissue lysate 10 ug
Lane 2: ab246486 IP in Human testis tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab246486 in human testis tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ly-6K antibody [EPR23021-61] (ab246486)](https://www.abcam.com/ps/products/246/ab246486/Images/ab246486-2-anti-ly6k-antibody-epr2302161-immunohistochemistry-human-heart.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ly-6K antibody [EPR23021-61] (ab246486)Immunohistochemical analysis of paraffin-embedded Human heart tissue labeling Ly-6K with ab246486 at 1/2000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Negative control: No staining in human heart (PMID:18089789). The section was incubated with ab246486 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 a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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![Anti-Ly-6K antibody [EPR23021-61] (ab246486)](https://www.abcam.com/ps/products/246/ab246486/Images/ab246486-5-benefits-of-recombinant-antibodies.png)
Anti-Ly-6K antibody [EPR23021-61] (ab246486)
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
Certificate of Compliance
References (0)
ab246486 has not yet been referenced specifically in any publications.
