Recombinant Anti-Annexin-6/ANXA6 antibody [EPR19517] KO Tested (ab201023)

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Western blot - 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

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.
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
For more information see here.
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.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19517
Isotype

IgG
Research areas

Alternative Versions
- Anti-Annexin-6/ANXA6 antibody [EPR19517] - BSA and Azide free (ab240359)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human ANXA6 (Annexin-6) knockout HeLa cell line (ab265677)
KO cell lysates
- Human ANXA6 (Annexin-6) knockout HeLa cell lysate (ab257351)
Recombinant Protein
- Recombinant Human Annexin-6/ANXA6 protein (ab92968)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

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
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.

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
modifications

Phosphorylated in response to growth factor stimulation.
Cellular localization

Cytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
Target information above from: UniProt accession P08133 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
- Unigene: 265347 Mouse
- Unigene: 204053 Rat
see all
Alternative names
- 67 kDa calelectrin antibody
- Annexin A6 antibody
- Annexin VI antibody
- Annexin VI p68 antibody
- Annexin-6 antibody
- AnnexinA6 antibody
- AnnexinVI antibody
- ANX 6 antibody
- ANX A6 antibody
- ANX6 antibody
- ANXA 6 antibody
- ANXA6 antibody
- ANXA6_HUMAN antibody
- Calcium binding protein p68 antibody
- Calelectrin antibody
- Calphobindin II antibody
- Calphobindin-II antibody
- CalphobindinII antibody
- CBP 68 antibody
- CBP68 antibody
- Chromobindin 20 antibody
- Chromobindin-20 antibody
- Chromobindin20 antibody
- CPB II antibody
- CPB-II antibody
- CPBII antibody
- Lipocortin VI antibody
- LipocortinVI antibody
- p68 antibody
- p70 antibody
- Protein III antibody
- ProteinIII antibody
see all

Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)

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.
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.
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.
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.
Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)

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 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

There was an additional band detected in the heart lysate.
Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)

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 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)

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.
Immunoprecipitation - Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)

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.
Anti-Annexin-6/ANXA6 antibody [EPR19517] (ab201023)

Western blot protocols
Immunoprecipitation protocols
Immunohistochemistry protocols

Click here to view the general protocols

Datasheet

Publishing research using ab201023? Please let us know so that we can cite the reference in this datasheet.

ab201023 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Recombinant Protein

Related Products

Applications

Target

Function

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Certificate of Compliance

References (0)

Customer reviews and Q&As

Post-translational
modifications