Anti-DNAJC3 antibody (ab70840)
Key features and details
- Rabbit polyclonal to DNAJC3
- Suitable for: IHC-P, ICC/IF, WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-DNAJC3 antibody
See all DNAJC3 primary antibodies -
Description
Rabbit polyclonal to DNAJC3 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, ICC/IF, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Chicken
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human DNAJC3.
Read Abcam's proprietary immunogen policy (Peptide available as ab72883.) -
Positive control
- This antibody gave a positive signal in the following human whole cell lysates: TE 671, Ramos, SK N BE, Hela - Staurosporine Treated (24hr, 500nM), Hela - Hydroxyurea Treated (48hr, 2uM), Jurkat - Staurosporine Treated (24hr, 500nM) and Y79.
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General notes
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab70840 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 Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. ICC/IF Use a concentration of 1 µg/ml. WB Use a concentration of 1 µg/ml. Detects a band of approximately 54 kDa (predicted molecular weight: 58 kDa). Target
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Function
Involved in the unfolded protein response (UPR) during ER stress. Co-chaperone of HSPA8/HSC70, it stimulates its ATPase activity. May inhibit both the autophosphorylation of EIF2AK2/PKR and the ability of EIF2AK2 to catalyze phosphorylation of the EIF2A. May inhibit EIF2AK3/PERK activity. -
Tissue specificity
Widely expressed with high level in the pancreas and testis. Also expressed in cell lines with different levels. -
Sequence similarities
Contains 1 J domain.
Contains 9 TPR repeats. -
Domain
The J domain mediates interaction with HSPA8. -
Cellular localization
Endoplasmic reticulum. - Information by UniProt
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Database links
- Entrez Gene: 5611 Human
- Entrez Gene: 100037258 Mouse
- Entrez Gene: 63880 Rat
- Omim: 601184 Human
- SwissProt: Q13217 Human
- SwissProt: Q91YW3 Mouse
- SwissProt: Q9R0T3 Rat
- Unigene: 59214 Human
see all -
Alternative names
- DnaJ (Hsp40) homolog subfamily C member 3 antibody
- DnaJ homolog subfamily C member 3 antibody
- DNAJC3 antibody
see all
Images
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Western blot - Anti-DNAJC3 antibody (ab70840)All lanes : Anti-DNAJC3 antibody (ab70840) at 1/250 dilution
Lane 1 : TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate
Lane 2 : Hela Whole Cell Lysate - Staurosporine Treated (24hr, 500nM)
Lane 3 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate
Lane 4 : Hela Whole Cell Lysate - Hydroxyurea Treated (48hr, 2uM)
Lane 5 : SK N BE (Human neuroblastoma) Whole Cell Lysate
Lane 6 : Jurkat Whole Cell Lysate - Staurosporine Treated (24hr, 500nM)
Lane 7 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 58 kDa
Observed band size: 54 kDa why is the actual band size different from the predicted? -
Immunocytochemistry/ Immunofluorescence - Anti-DNAJC3 antibody (ab70840)ICC/IF image of ab70840 stained HeLa cells. The cells were 100% Methanol fixed (5 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab70840, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% Methanol fixed (5 min) Hek293, HepG2, MCF-7 cells at 1µg/ml. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC3 antibody (ab70840)IHC image of ab70840 staining in human testis formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab70840, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Protocols
Datasheets and documents
References (1)
ab70840 has been referenced in 1 publication.
- Lin Y & Sun Z In vivo pancreatic ß-cell-specific expression of antiaging gene Klotho: a novel approach for preserving ß-cells in type 2 diabetes. Diabetes 64:1444-58 (2015). IHC . PubMed: 25377875
Images
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Western blot - Anti-DNAJC3 antibody (ab70840)All lanes : Anti-DNAJC3 antibody (ab70840) at 1/250 dilution
Lane 1 : TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate
Lane 2 : Hela Whole Cell Lysate - Staurosporine Treated (24hr, 500nM)
Lane 3 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate
Lane 4 : Hela Whole Cell Lysate - Hydroxyurea Treated (48hr, 2uM)
Lane 5 : SK N BE (Human neuroblastoma) Whole Cell Lysate
Lane 6 : Jurkat Whole Cell Lysate - Staurosporine Treated (24hr, 500nM)
Lane 7 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 58 kDa
Observed band size: 54 kDa why is the actual band size different from the predicted?
-
Immunocytochemistry/ Immunofluorescence - Anti-DNAJC3 antibody (ab70840)ICC/IF image of ab70840 stained HeLa cells. The cells were 100% Methanol fixed (5 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab70840, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% Methanol fixed (5 min) Hek293, HepG2, MCF-7 cells at 1µg/ml.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC3 antibody (ab70840)
IHC image of ab70840 staining in human testis formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab70840, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
