Recombinant Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)
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Overview
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Product name
Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free
See all Clusterin primary antibodies -
Description
Rabbit monoclonal [EPR2911] to Clusterin - Low endotoxin, Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IHC-FoFr, IHC-P, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
aa 400-500.
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Positive control
- IHC-P: human breast carcinoma and testis tissue; WB: human testis, brain, tonsil, fetal brain and plasma lysates
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General notes
ab229445 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.
Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.
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: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2911 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab229445 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-FoFr | Use at an assay dependent concentration. | |
| IHC-P | Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
| WB | Use at an assay dependent concentration. Predicted molecular weight: 52 kDa. |
Target
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Function
Isoform 1 functions as extracellular chaperone that prevents aggregation of nonnative proteins. Prevents stress-induced aggregation of blood plasma proteins. Inhibits formation of amyloid fibrils by APP, APOC2, B2M, CALCA, CSN3, SNCA and aggregation-prone LYZ variants (in vitro). Does not require ATP. Maintains partially unfolded proteins in a state appropriate for subsequent refolding by other chaperones, such as HSPA8/HSC70. Does not refold proteins by itself. Binding to cell surface receptors triggers internalization of the chaperone-client complex and subsequent lysosomal or proteasomal degradation. Secreted isoform 1 protects cells against apoptosis and against cytolysis by complement. Intracellular isoforms interact with ubiquitin and SCF (SKP1-CUL1-F-box protein) E3 ubiquitin-protein ligase complexes and promote the ubiquitination and subsequent proteasomal degradation of target proteins. Promotes proteasomal degradation of COMMD1 and IKBKB. Modulates NF-kappa-B transcriptional activity. Nuclear isoforms promote apoptosis. Mitochondrial isoforms suppress BAX-dependent release of cytochrome c into the cytoplasm and inhibit apoptosis. Plays a role in the regulation of cell proliferation. -
Tissue specificity
Detected in blood plasma, cerebrospinal fluid, milk, seminal plasma and colon mucosa. Detected in the germinal center of colon lymphoid nodules and in colon parasympathetic ganglia of the Auerbach plexus (at protein level). Ubiquitous. Detected in brain, testis, ovary, liver and pancreas, and at lower levels in kidney, heart, spleen and lung. -
Sequence similarities
Belongs to the clusterin family. -
Post-translational
modificationsIsoform 1 is proteolytically cleaved on its way through the secretory system, probably within the Golgi lumen.
Polyubiquitinated, leading to proteasomal degradation.
Heavily N-glycosylated. About 30% of the protein mass is comprised of complex N-linked carbohydrate. -
Cellular localization
Secreted. Can retrotranslocate from the secretory compartments to the cytosol upon cellular stress and Nucleus. Cytoplasm. Mitochondrion membrane. Cytoplasm, cytosol. Microsome. Endoplasmic reticulum. Cytoplasmic vesicle, secretory vesicle, chromaffin granule. Isoforms lacking the N-terminal signal sequence have been shown to be cytoplasmic and/or nuclear. Secreted isoforms can retrotranslocate from the secretory compartments to the cytosol upon cellular stress. Detected in perinuclear foci that may be aggresomes containing misfolded, ubiquitinated proteins. Detected at the mitochondrion membrane upon induction of apoptosis. - Information by UniProt
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Database links
- Entrez Gene: 1191 Human
- Omim: 185430 Human
- SwissProt: P10909 Human
- Unigene: 436657 Human
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Alternative names
- 40 antibody
- AAG 4 antibody
- AAG4 antibody
see all
Images
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![Western blot - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)](https://www.abcam.com/ps/products/229/ab229445/Images/ab229445-317318-ab92548KOWBGDB.jpg)
Western blot - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)All lanes : Anti-Clusterin antibody [EPR2911] (ab92548) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Clusterin knockout HAP1 whole cell lysate
Lane 3 : U87-MG whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 52 kDaLanes 1 - 3: Merged signal (red and green). Green - ab92548 observed at 68 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab92548 was shown to recognize Clusterin in wild-type HAP1 cells as signal was lost at the expected MW in Clusterin knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Clusterin knockout samples were subjected to SDS-PAGE. Ab92548 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229445). -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)](https://www.abcam.com/ps/products/229/ab229445/Images/ab229445-4-anti-clusterin-antibody-epr2911-immunohistochemistry-breast-carcinoma-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling Clusterin with purified ab92548 at 1/450 dilution (0.23 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92548)
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![Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)](https://www.abcam.com/ps/products/229/ab229445/Images/ab229445-295382-ab92548IHCFr.jpg)
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)This IHC data was generated using the same anti-Clusterin antibody clone, EPR2911, in a different buffer formulation (cat# ab925480).
ab92548 (unpurified) staining Apolipoprotein J in mouse brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde, cut into 20 micron slices, permeablized with 0.1 M PBS with 1% Triton X and blocked with 10% serum for 60 minutes at 24°C. The sample was incubated with primary antibody (1/100 in 0.1M PBST with 10% donkeys serum) at 4°C for 24 hours. An Alexa Fluor® 488-conjugated donkey polyclonal (1/1000) was used as the secondary antibody.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)](https://www.abcam.com/ps/products/229/ab229445/Images/ab229445-295381-ab92548IHC.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)This IHC data was generated using the same anti-Clusterin antibody clone, EPR2911, in a different buffer formulation (cat# ab925480).
ab92548 (unpurified) at 1/100 dilution staining Apolipoprotein J in Human testis tissue by Immunohistochemistry Formalin-fixed, Paraffin-embedded tissue.
Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
Datasheets and documents
References
ab229445 has not yet been referenced specifically in any publications.
