Recombinant Anti-Annexin-6/ANXA6 antibody [EPR17308] - N-terminal (ab199422)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17308] to Annexin-6/ANXA6 - N-terminal
- Suitable for: IP, ICC/IF, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Annexin-6/ANXA6 antibody [EPR17308] - N-terminal
See all Annexin-6/ANXA6 primary antibodies -
Description
Rabbit monoclonal [EPR17308] to Annexin-6/ANXA6 - N-terminal -
Host species
Rabbit -
Tested applications
Suitable for: IP, ICC/IF, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment within Human Annexin-6/ANXA6 aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: P08133 -
Positive control
- WB: Raji, HeLa and Jurkat whole cell lysate. Mouse brain, kidney and spleen tissue lysate. Rat brain, heart, kidney and spleen tissue lysate. ICC/IF: HeLa and K562 cells.
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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
EPR17308 -
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
Applications
Our Abpromise guarantee covers the use of ab199422 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 |
---|---|---|
IP | Use at an assay dependent concentration. | |
ICC/IF | 1/100. | |
WB | 1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 76 kDa). |
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 [EPR17308] - N-terminal (ab199422) 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.
Performed under reducing conditions.
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 - ab199422 observed at 75 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab199422 was shown to react with Annexin-6/ANXA6 in HeLa wild-type cells in western blot with loss of signal observed in ANXA6 knockout cell line ab265677 (ANXA6 knockout cell lysate ab257351). Wild-type and ANXA6 knockout HeLa cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab199422 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-Annexin-6/ANXA6 antibody [EPR17308] - N-terminal (ab199422)
Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling Annexin-6/ANXA6 with ab199422 at 1/100, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 (green). Cytoplasm staining on K562 cell line is observed (Subcellular Location: Cytoplasm [UniProt]). The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 (red).
The negative controls are as follows:-
-ve control 1 - ab199422 at 1/1000 followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500.
-ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500. -
All lanes : Anti-Annexin-6/ANXA6 antibody [EPR17308] - N-terminal (ab199422) at 1/10000 dilution
Lane 1 : Raji (Human Burkitt's lymphoma) whole cell lysate
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma ) whole cell lysate
Lane 3 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 76 kDa
Observed band size: 68 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking and diluting buffer was 5% NFDM/TBST.
The observed MW is consistent with what has been described in the literature (PMID:12140262).
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All lanes : Anti-Annexin-6/ANXA6 antibody [EPR17308] - N-terminal (ab199422) at 1/1000 dilution
Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse kidney tissue lysate
Lane 3 : Mouse spleen tissue lysate
Lane 4 : Rat brain tissue lysate
Lane 5 : Rat heart tissue lysate
Lane 6 : Rat kidney tissue lysate
Lane 7 : Rat spleen tissue lysate
Lane 8 : PC-12 (Rat adrenal gland pheochromocytoma) cell lysate
Lane 9 : NIH/3T3 (Mouse embryo fibroblast) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 76 kDa
Observed band size: 68 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking and diluting buffer was 5% NFDM/TBST.
The observed MW is consistent with what has been described in the literature (PMID:12140262). -
Immunocytochemistry/ Immunofluorescence - Anti-Annexin-6/ANXA6 antibody [EPR17308] - N-terminal (ab199422)
Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Annexin-6/ANXA6 with ab199422 at 1/100, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 (green). Cytoplasm staining on HeLa cell line is observed (Subcellular Location: Cytoplasm [UniProt]). The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 (red).
The negative controls are as follows:-
-ve control 1 - ab199422 at 1/1000 followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500.
-ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500. -
Annexin-6/ANXA6 was immunoprecipitated from 1mg of Raji (Human Burkitt's lymphoma cell line) whole cell extract with ab199422 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab199422 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: Raji whole cell extract 10 µg (Input).
Lane 2: ab199422 IP in Raji whole cell extract.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab199422 in Raji whole cell extract.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Protocols
Datasheets and documents
Certificate of Compliance
References (0)
ab199422 has not yet been referenced specifically in any publications.