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
Related conjugates and formulations
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. This information is proprietary to Abcam and/or its suppliers.
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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 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: 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
The Abpromise guarantee
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.
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WB |
1/1000. Detects a band of approximately 72 kDa (predicted molecular weight: 76 kDa).
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IP |
1/40.
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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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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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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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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
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab201023 has not yet been referenced specifically in any publications.