Recombinant Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19517] to Annexin-6/ANXA6
- Suitable for: IHC-P, WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Annexin-6/ANXA6 antibody [EPR19517]
See all Annexin-6/ANXA6 primary antibodies -
Description
Rabbit monoclonal [EPR19517] to Annexin-6/ANXA6 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment within Human Annexin-6/ANXA6 aa 200-550. The exact sequence is proprietary.
Database link: P08133 -
Positive control
- WB: Human fetal liver, fetal heart, fetal kidney and fetal spleen lysates; A549, HeLa, Jurkat, Raji, C6, NIH/3T3 and PC-12 whole cell lysates; Mouse brain, heart, spleen and kidney lysates; Rat brain, heart, kidney and spleen lysates. IHC-P: Human tonsil, pancreas and stomach tissues. IP: Human fetal liver whole cell lysate.
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General notes
This product was previously labelled as Annexin VI
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19517 -
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
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Recombinant Protein
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Related Products
Applications
Our Abpromise guarantee covers the use of ab201023 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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IHC-P | 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. | |
WB | 1/1000. Detects a band of approximately 72 kDa (predicted molecular weight: 76 kDa). | |
IP | 1/40. |
Target
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Function
May associate with CD21. May regulate the release of Ca(2+) from intracellular stores. -
Sequence similarities
Belongs to the annexin family.
Contains 8 annexin repeats. -
Domain
A pair of annexin repeats may form one binding site for calcium and phospholipid. -
Post-translational
modificationsPhosphorylated in response to growth factor stimulation. -
Cellular localization
Cytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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Database links
- Entrez Gene: 309 Human
- Entrez Gene: 11749 Mouse
- Entrez Gene: 79125 Rat
- Omim: 114070 Human
- SwissProt: P08133 Human
- SwissProt: P14824 Mouse
- SwissProt: P48037 Rat
- Unigene: 412117 Human
see all -
Alternative names
- 67 kDa calelectrin antibody
- Annexin A6 antibody
- Annexin VI antibody
see all
Images
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All lanes : Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ANXA6 knockout HeLa cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : Raji 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: 76 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab201023 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa.
ab201023 Anti-Annexin-6/ANXA6 antibody [EPR19536] was shown to specifically react with 6/ANXA6 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265677 (knockout cell lysate ab257351) was used. Wild-type and 6/ANXA6 knockout samples were subjected to SDS-PAGE. ab201023 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Annexin-6/ANXA6 with ab201023 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Cytoplasm staining on lymphocytes of Human tonsil is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling Annexin-6/ANXA6 with ab201023 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Membrane staining on Human pancreas islet is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)
Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling Annexin-6/ANXA6 with ab201023 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Cytoplasm staining on stromal cells of Human stomach is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023) at 1/5000 dilution
Lane 1 : Human fetal liver lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lane 4 : Human fetal spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 76 kDa
Observed band size: 72 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
There was an additional band detected in the heart lysate.
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All lanes : Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023) at 1/1000 dilution
Lane 1 : A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 4 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 76 kDa
Observed band size: 72 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023) at 1/1000 dilution
Lane 1 : Mouse brain lysate at 10 µg
Lane 2 : Mouse heart lysate at 10 µg
Lane 3 : Mouse spleen lysate at 10 µg
Lane 4 : Mouse kidney lysate at 10 µg
Lane 5 : Rat brain lysate at 10 µg
Lane 6 : Rat heart lysate at 10 µg
Lane 7 : Rat kidney lysate at 10 µg
Lane 8 : Rat spleen lysate at 10 µg
Lane 9 : C6 (Rat glial tumor cell line) whole cell lysate at 20 µg
Lane 10 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg
Lane 11 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 76 kDa
Observed band size: 72 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1,2 and 3: 15 seconds; Lane 4: 3 minutes; Lane 5,6,7 and 8: 15 seconds; Lane 9 and 10: 5 seconds; Lane 11: 3 minutes.
There was an additional band detected in the heart lysate.
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Annexin-6/ANXA6 was immunoprecipitated from 1mg of Human fetal liver whole cell lysate with ab201023 at 1/40 dilution.
Western blot was performed from the immunoprecipitate using ab201023 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: Human fetal liver whole cell lysate, 10µg (Input).
Lane 2: ab201023 IP in Human fetal liver whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control - (ab172730) instead of ab201023 in Human fetal liver whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
Protocols
Datasheets and documents
Certificate of Compliance
References (0)
ab201023 has not yet been referenced specifically in any publications.