Recombinant Anti-Eph receptor B4/HTK antibody [EPR23221-54] - BSA and Azide free (ab269867)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23221-54] to Eph receptor B4/HTK - BSA and Azide free
- Suitable for: IP, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Eph receptor B4/HTK antibody [EPR23221-54] - BSA and Azide free
See all Eph receptor B4/HTK primary antibodies -
Description
Rabbit monoclonal [EPR23221-54] to Eph receptor B4/HTK - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IP, WBmore details
Unsuitable for: Flow Cyt,ICC/IF or IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK293T, HT-29 and MCF7 lysates. IP: HT-29 and HUVEC whole cell lysates.
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General notes
ab269867 is the carrier-free version of ab254300.
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
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. 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
EPR23221-54 -
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 ab269867 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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IP |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 108 kDa.
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Notes |
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IP
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 108 kDa. |
Target
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Function
Receptor for members of the ephrin-B family. Binds to ephrin-B2. May have a role in events mediating differentiation and development. -
Tissue specificity
Abundantly expressed in placenta and in a range of primary tissues and malignant cell lines. Expressed in fetal, but not adult, brain, and in primitive and myeloid, but not lymphoid, hematopoietic cells. -
Sequence similarities
Belongs to the protein kinase superfamily. Tyr protein kinase family. Ephrin receptor subfamily.
Contains 2 fibronectin type-III domains.
Contains 1 protein kinase domain.
Contains 1 SAM (sterile alpha motif) domain. -
Post-translational
modificationsAutophosphorylated. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 2050 Human
- Omim: 600011 Human
- SwissProt: P54760 Human
- Unigene: 437008 Human
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Alternative names
- Ephb4 antibody
- EPHB4_HUMAN antibody
- Ephrin receptor EphB4 antibody
see all
Images
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All lanes : Anti-Eph receptor B4/HTK antibody [EPR23221-54] (ab254300) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : EPHB4 knockout HEK293T cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : HT-29 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: 108 kDa
Observed band size: 125 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab254300).
Lanes 1-4: Merged signal (red and green). Green - ab254300 observed at 125 kDa. Red - loading control ab8245 observed at 36 kDa.
ab254300 Anti-Eph receptor B4/HTK antibody [EPR23221-54] was shown to specifically react with Eph receptor B4/HTK in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266733 (knockout cell lysate ab257429) was used. Wild-type and Eph receptor B4/HTK knockout samples were subjected to SDS-PAGE. ab254300 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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Eph receptor B4 was immunoprecipitated from 0.35 mg HUVEC (human umbilical vein endothelial cell) whole cell lysate with ab254300 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254300 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Lane 1: HUVEC (human umbilical vein endothelial cell) whole cell lysate 10ug
Lane 2: ab254300 IP in HUVEC whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab254300 in HUVEC whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds.
Cleaved intracellular domain (ICD) was observed at around 47kDa. (PMID: 24854540).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254300).
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Eph receptor B4 was immunoprecipitated from 0.35 mg HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate with ab254300 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254300 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Lane 1: HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2: ab254300 IP in HT-29 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab254300 in HT-29 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds.
Cleaved intracellular domain (ICD) was observed at around 47kDa. (PMID: 24854540).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254300).
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)
ab269867 has not yet been referenced specifically in any publications.