Recombinant Anti-Clusterin beta chain antibody [EPR17538-101] - BSA and Azide free (ab229127)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17538-101] to Clusterin beta chain - BSA and Azide free
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Mouse, Rat
Related conjugates and formulations
Overview
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Product name
Anti-Clusterin beta chain antibody [EPR17538-101] - BSA and Azide free
See all Clusterin beta chain primary antibodies -
Description
Rabbit monoclonal [EPR17538-101] to Clusterin beta chain - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat -
Immunogen
Recombinant fragment aa 1-250. The exact sequence is proprietary.
Database link: Q06890 -
Positive control
- IHC-P: Mouse spleen tissue.
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General notes
ab229127 is the carrier-free version of ab184099.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17538-101 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab229127 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 |
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 52 kDa.
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Notes |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 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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Alternative names
- 40 antibody
- Aging-associated gene 4 protein antibody
- Apo-J antibody
see all
Images
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This data was developed using ab184099, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded (A) Testis tissue from wild-type C57BL/6J mice (B) Testis tissue from Clusterin knockout mice staining with ab184099 at 1/2000 dilution and ready-to-use Goat Anti-Rabbit IgG H&L (HRP) secondary. Counterstaining with hematoxylin. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins. Positive staining on (A) Testis tissue from wild-type C57BL/6J mice and no staining on (B) Testis tissue from Clusterin knockout mice. The section was incubated with ab184099 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND™ RX instrument. The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Clusterin-KO homozygous mice (Strain ID: T014419). -
This data was developed using ab184099, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded (A) Spleen tissue from wild-type C57BL/6J mice (B) Spleen tissue from Clusterin knockout mice staining with ab184099 at 1/2000 dilution and ready-to-use Goat Anti-Rabbit IgG H&L (HRP) secondary. Counterstaining with hematoxylin. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins. Positive staining on (A) Spleen tissue from wild-type C57BL/6J mice and no staining on (B) Spleen tissue from Clusterin knockout mice. The section was incubated with ab184099 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND™ RX instrument.
The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Clusterin-KO homozygous mice (Strain ID: T014419). -
This data was developed using ab184099, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded (A) Adrenal gland tissue from wild-type C57BL/6J mice (B) Adrenal gland tissue from Clusterin knockout mice staining with ab184099 at 1/2000 dilution and ready-to-use Goat Anti-Rabbit IgG H&L (HRP) secondary. Counterstaining with hematoxylin. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins. Positive staining on (A) Adrenal gland tissue from wild-type C57BL/6J mice and no staining on (B) Adrenal gland tissue from Clusterin knockout mice. The section was incubated with ab184099 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND™ RX instrument.The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Clusterin-KO homozygous mice (Strain ID: T014419). -
All lanes : Anti-Clusterin beta chain antibody [EPR17538-101] (ab184099) at 1/1000 dilution
Lane 1 : Wild-type mouse adrenal gland tissue lysate (male)
Lane 2 : Clusterin knockout adrenal gland tissue lysate (male)
Lane 3 : Wild-type mouse spleen tissue lysate (male)
Lane 4 : Clusterin knockout spleen tissue lysate (male)
Lane 5 : Wild-type mouse testis tissue lysate (male)
Lane 6 : Clusterin knockout testis tissue lysate (male)
Lane 7 : Wild-type mouse adrenal gland tissue lysate (female)
Lane 8 : Clusterin knockout adrenal gland tissue lysate (female)
Lane 9 : Wild-type mouse spleen tissue lysate (female)
Lane 10 : Clusterin knockout spleen tissue lysate (female)
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 52 kDa
Observed band size: 33 kDa why is the actual band size different from the predicted?
Exposure time: 10 seconds
This data was developed using ab184099, the same antibody clone in a different buffer formulation.
The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Clusterin-KO homozygous mice (Strain ID: T014419). -
Immunohistochemical analysis of paraffin-embedded rat adrenal gland tissue labeling Clusterin with ab184099 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Stronger cytoplasmic staining in the medulla than the cortex of rat adrenal gland (PMID: 16436671) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184099).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Clusterin with ab184099 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in stromal cells of mouse spleen (PMID: 24865838) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184099).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-Clusterin beta chain antibody [EPR17538-101] - BSA and Azide free (ab229127) at 1/1000 dilution
Lane 1 : Mouse lung tissue lysate
Lane 2 : Mouse adrenal gland tissue lysate
Lane 3 : Rat adrenal gland tissue lysate
Lane 4 : Rat lung tissue lysate
Lane 5 : Mouse stomach tissue lysate
Lane 6 : Rat stomach tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 52 kDa
Observed band size: 33 kDa why is the actual band size different from the predicted?
Additional bands at: 70 kDa (possible immature (unprocessed))Blocking and dilution buffer: 5% NFDM/TBST .
Exposure times.
Lanes 1, 2 & 3: 3 minutes.
Lane 4: 53 seconds.
Lanes 5 & 6: 100 seconds.
The molecular masses observed are consistent with what has been described in the literature (PMID: 8373947).
The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab229127 has not yet been referenced specifically in any publications.