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Western Blot (WB) (Figure 1. Western blot analysis of DNAJC6 using anti-DNAJC6 antibody (MBS1753704).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: rat brain tissue lysatesLane 2: mouse brain tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-DNAJC6 antigen affinity purified polyclonal antibody (Catalog # MBS1753704) at 0. 25 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for DNAJC6 at approximately 120KD. The expected band size for DNAJC6 is at 120KD. )

Rabbit DNAJC6 Polyclonal Antibody | anti-DNAJC6 antibody

Anti-DNAJC6 Antibody

Gene Names
DNAJC6; DJC6; PARK19
Reactivity
Human, Mouse, Rat
Applications
Western Blot, Immunohistochemistry, Immunocytochemistry, Immunofluorescence, Flow Cytometry, Functional Assay, ELISA
Purity
Immunogen affinity purified.
Synonyms
DNAJC6; Polyclonal Antibody; Anti-DNAJC6 Antibody; Dnajc6; Kiaa0473; Putative tyrosine-protein phosphatase auxilin; DnaJ homolog subfamily C member 6; DnaJ heat shock protein family (Hsp40) member C6; anti-DNAJC6 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Mouse, Rat
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Rabbit IgG polyclonal antibody for DNAJC6 detection.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Applicable Applications for anti-DNAJC6 antibody
Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA)
Application Notes
WB: 0.1-0.25ug/ml|Mouse, Rat|
IHC-P: 2-5ug/ml|Human, Mouse|
ICC/IF: 5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Direct ELISA: 0.1-0.5ug/ml|Human|
Immunogen
E Coli-derived human DNAJC6 recombinant protein (Position: R32-Y913).
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Recommended Detection Systems
Recommended Detection Systems
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

Western Blot (WB)

(Figure 1. Western blot analysis of DNAJC6 using anti-DNAJC6 antibody (MBS1753704).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: rat brain tissue lysatesLane 2: mouse brain tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-DNAJC6 antigen affinity purified polyclonal antibody (Catalog # MBS1753704) at 0. 25 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for DNAJC6 at approximately 120KD. The expected band size for DNAJC6 is at 120KD. )

Western Blot (WB) (Figure 1. Western blot analysis of DNAJC6 using anti-DNAJC6 antibody (MBS1753704).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: rat brain tissue lysatesLane 2: mouse brain tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-DNAJC6 antigen affinity purified polyclonal antibody (Catalog # MBS1753704) at 0. 25 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for DNAJC6 at approximately 120KD. The expected band size for DNAJC6 is at 120KD. )

Immunohistochemistry (IHC)

(Figure 2. IHC analysis of DNAJC6 using anti-DNAJC6 antibody (MBS1753704).DNAJC6 was detected in paraffin-embedded section of human meningeoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-DNAJC6 Antibody (MBS1753704) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )

Immunohistochemistry (IHC) (Figure 2. IHC analysis of DNAJC6 using anti-DNAJC6 antibody (MBS1753704).DNAJC6 was detected in paraffin-embedded section of human meningeoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-DNAJC6 Antibody (MBS1753704) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )

Immunohistochemistry (IHC)

(Figure 3. IHC analysis of DNAJC6 using anti-DNAJC6 antibody (MBS1753704).DNAJC6 was detected in paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-DNAJC6 Antibody (MBS1753704) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )

Immunohistochemistry (IHC) (Figure 3. IHC analysis of DNAJC6 using anti-DNAJC6 antibody (MBS1753704).DNAJC6 was detected in paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-DNAJC6 Antibody (MBS1753704) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )

Immunofluorescence (IF)

(Figure 4. IF analysis of DNAJC6 using anti- DNAJC6 antibody (MBS1753704).DNAJC6 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-DNAJC6 Antibody (MBS1753704) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )

Immunofluorescence (IF) (Figure 4. IF analysis of DNAJC6 using anti- DNAJC6 antibody (MBS1753704).DNAJC6 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-DNAJC6 Antibody (MBS1753704) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )

Flow Cytometry (FC/FACS)

(Figure 5. Flow Cytometry analysis of SiHa cells using anti-DNAJC6 antibody (MBS1753704).Overlay histogram showing SiHa cells stained with MBS1753704 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DNAJC6 Antibody (MBS1753704, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Flow Cytometry (FC/FACS) (Figure 5. Flow Cytometry analysis of SiHa cells using anti-DNAJC6 antibody (MBS1753704).Overlay histogram showing SiHa cells stained with MBS1753704 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DNAJC6 Antibody (MBS1753704, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Related Product Information for anti-DNAJC6 antibody
Putative tyrosine-protein phosphatase auxilin is an enzyme that in humans is encoded by the DNAJC6 gene. DNAJC6 belongs to the evolutionarily conserved DNAJ/HSP40 family of proteins, which regulate molecular chaperone activity by stimulating ATPase activity. DNAJ proteins may have up to 3 distinct domains: a conserved 70-amino acid J domain, usually at the N terminus, a glycine/phenylalanine (G/F)-rich region, and a cysteine-rich domain containing 4 motifs resembling a zinc finger domain.
References
1. Edvardson, S., Cinnamon, Y., Ta-Shma, A., Shaag, A., Yim, Y. -I., Zenvirt, S., Jalas, C., Lesage, S., Brice, A., Taraboulos, A., Kaestner, K. H., Greene, L. E., Elpeleg, O. A deleterious mutation in DNAJC6 encoding the neuronal-specific clathrin-uncoating co-chaperone auxilin, is associated with juvenile parkinsonism. PLoS One 7: e36458, 2012. Note: Electronic Article.
2. Elsayed, L. E. O., Drouet, V., Usenko, T., Mohammed, I. N., Hamed, A. A. A., Elseed, M. A., Salih, M. A. M., Koko, M. E., Mohamed, A. Y. O., Siddig, R. A., Elbashir, M. I., Ibrahim, M. E., Durr, A., Stevanin, G., Lesage, S., Ahmed, A. E., Brice, A. A novel nonsense mutation in DNAJC6 expands the phenotype of autosomal-recessive juvenile-onset Parkinson's disease. (Letter) Ann. Neurol. 79: 335-338, 2016.
3. Fotin, A., Cheng, Y., Grigorieff, N., Walz, T., Harrison, S. C., Kirchhausen, T. Structure of an auxilin-bound clathrin coat and its implications for the mechanism of uncoating. Nature 432: 649-653, 2004.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
99,997 Da
NCBI Official Full Name
putative tyrosine-protein phosphatase auxilin isoform 1
NCBI Official Synonym Full Names
DnaJ (Hsp40) homolog, subfamily C, member 6
NCBI Official Symbol
DNAJC6
NCBI Official Synonym Symbols
DJC6; PARK19
NCBI Protein Information
putative tyrosine-protein phosphatase auxilin; DnaJ (Hsp40) homolog, subfamily B, member 6
UniProt Protein Name
Putative tyrosine-protein phosphatase auxilin
UniProt Gene Name
DNAJC6
UniProt Synonym Gene Names
KIAA0473
UniProt Entry Name
AUXI_HUMAN

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Product Notes

The DNAJC6 dnajc6 (Catalog #AAA1753704) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-DNAJC6 Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's DNAJC6 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.1-0.25ug/ml|Mouse, Rat| IHC-P: 2-5ug/ml|Human, Mouse| ICC/IF: 5ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human| Direct ELISA: 0.1-0.5ug/ml|Human|. Researchers should empirically determine the suitability of the DNAJC6 dnajc6 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "DNAJC6, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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