Overview

  • Product name
    Anti-Clusterin antibody [CLI-9]
    See all Clusterin primary antibodies
  • Description
    Mouse Monoclonal [CLI-9] to Clusterin
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, IHC-P, ICC, IHC-Fr, IP, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Full length native protein (purified) corresponding to Human Clusterin .

Properties

Applications

Our Abpromise guarantee covers the use of ab16077 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
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

IHC-P Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB 1/2000. Predicted molecular weight: 54 kDa.

Target

  • 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
    modifications
    Isoform 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
  • Database links
  • Alternative names
    • 40 antibody
    • AAG 4 antibody
    • AAG4 antibody
    • Aging associated protein 4 antibody
    • Aging-associated gene 4 protein antibody
    • AI893575 antibody
    • APO J antibody
    • Apo-J antibody
    • APOJ antibody
    • ApoJalpha antibody
    • ApoJbeta antibody
    • Apolipoprotein J antibody
    • ApolipoproteinJ antibody
    • CLI antibody
    • CLU antibody
    • CLU1 antibody
    • CLU2 antibody
    • CLUS_HUMAN antibody
    • Clusterin alpha chain antibody
    • Clusterin antibody
    • Clusterin beta chain antibody
    • Complement associated protein SP 40 40 antibody
    • Complement associated protein SP 40 antibody
    • Complement associated protein SP40 antibody
    • Complement cytolysis inhibitor a chain antibody
    • Complement cytolysis inhibitor antibody
    • Complement cytolysis inhibitor b chain antibody
    • Complement lysis inhibitor antibody
    • Complement-associated protein SP-40 antibody
    • D14Ucla3 antibody
    • Dimeric acid glycoprotein antibody
    • Glycoprotein 80 antibody
    • Glycoprotein III antibody
    • GP80 antibody
    • Ku70-binding protein 1 antibody
    • KUB 1 antibody
    • KUB1 antibody
    • MGC24903 antibody
    • NA1/NA2 antibody
    • RATTRPM2B antibody
    • SGP 2 antibody
    • SGP2 antibody
    • SP 40 antibody
    • SP40 antibody
    • Sugp-2 antibody
    • Sulfated glycoprotein 2 antibody
    • Testosterone repressed prostate message 2 antibody
    • Testosterone-repressed prostate message 2 antibody
    • TRPM 2 antibody
    • TRPM-2 antibody
    • TRPM2 antibody
    • TRPM2B antibody
    • Trpmb antibody
    see all

Images

  • Overlay histogram showing HeLa cells stained with ab16077 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16077, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

This product has been referenced in:
  • Peng L  et al. Serum proteomics analysis and comparisons using iTRAQ in the progression of hepatitis B. Exp Ther Med 6:1169-1176 (2013). WB ; Human . Read more (PubMed: 24223640) »
  • Sinha R  et al. Selenium-Responsive Proteins in the Sera of Selenium-Enriched Yeast-Supplemented Healthy African American and Caucasian Men. Cancer Epidemiol Biomarkers Prev : (2010). WB ; Human . Read more (PubMed: 20643827) »
See all 5 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Answer

Thank you for your enquiry.



I am glad to hear that another product has worked for you. However, it does mean that the ab16077 that you purchased earlier may be faulty. In which case, I am more than happy to offer you a free primary antibody of your choice from our catalogue. Please provide the catalogue number and your order details (sub-dealer company name, person in charge name, order date, lot number) so that I can trace your order and arrange for a replacement product to you.



As a formality, could you please fill up the WB questionnaire form attached and provide as much protocol information as possible? I have attached one English and one Japanese, and you only need to fill up either one. With the information you provide, we will be able to record the problem with ab16077 and retest the condition/lot/product if necessary.



Should you wish to get a refund instead, could you please contact your local sub-dealer which you purchased the product from? I am afraid that for refunds, it is between your organization and the sub-dealer to process and I do not have authority on this.



Please let me know how you would like to proceed with this matter.



Thank you.

Read More

Answer

Thank you for getting back to me.



You are correct that "reducing condition" means adding the BME or DTT so that the S-S bonds are cleaved. However, "denaturing condition" actually means boiling, where the proteinthree-dimensional structure will unfold and the epitope is exposed for antibody detection. Since you have performed reducing condition plus boiling, then I don't think that is a problem.I have attached our WB Protocol Guide link below for your reference.


https://www.abcam.com/index.html?pageconfig=resource&rid=11379#A6



Regarding the amount of protein, it is common to first measure the amount of sample protein so that you can compare whether the difference in band detection is based on sample treatment, and not because you have added different amounts of protein in each lane.



Did you find the publication from Jones and Jomary2002, helpful? Is there anything that I cando for you regarding this case?



I hope I can help you resolve the matter, so please feel free to contact me again if you have any otherquestions.

Read More

Answer

Thank you for your enquiry.



Firstly, regarding the expected molecular weight (54kDa) stated on the datasheet, the information was taken from SwissProt database (http://www.uniprot.org/uniprot/P10909). As you may already know, the protein has quite a few glycosylation sites andmay be the reason why the detected/reported band sizes are dissimilar from the SwissProt molecular weight.



Looking at your WB results, I think the data you obtained is correct. The publication you referred to also did mentioned that a ˜70kDa is the disulfied-linked alpha and beta subunit, in other words, unprocessed glycoprotein. We had 2 abreview feedbacks from customers who used denaturing condition (heating of sample before applying to SDS-PAGE) and obtained the alpha and beta subunits (˜40kDa). Can I confirm that your reducing condition also means that you have denatured the sample?If not, please try with denaturing condition.



Abreviews showing detection of 40kDa band (alpha, beta subunit)


https://www.abcam.co.jp/index.html?datasheet=16077&tab=abreviews&intabreviewid=6848



https://www.abcam.co.jp/index.html?datasheet=16077&tab=abreviews&intabreviewid=10815



Out of curiosity, how much protein (ug) did you load onto your gel? Normally, we would recommend around 20-40ug per well and if the volume is greater than this, there is a possibility that multimers may form causing band sizes to be larger than expected.



Also, one of the customers did provide areference regarding the protein and I hope this will be helpful to you. Unfortunately, I do not have access to the journal from my end but I am sure your university library should have a copy of the publication in archieve.



Jones and Jomary. Clusterin. Int J Biochem Cell Biol. 2002 May;34(5):427-31.

http://www.ncbi.nlm.nih.gov/pubmed/11906815



I hope the information above will be useful. If you have any other questions please do not hesitate to contact me.



Have a nice day!

Read More

Answer

That is super news. I am really pleased the alternative product worked well. As you have the images available I wonder whether you would like to submit an Abreview for ab69644? We currently have no Abreviews so will be eligible for extra points. Abreviews can be submitted by heading to the Abreviews tab on the product datasheet. We just need some basic protocol information. Please do not hesitate to contact us should you need further help in the future. All the best in your future research.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Human Brain, parafin embedded)
Specification
Human Brain, parafin embedded
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted May 19 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Tissue lysate - other (Human Plasma and Follicular Fluid)
Loading amount
18 µg
Specification
Human Plasma and Follicular Fluid
Gel Running Conditions
Reduced Denaturing (12% gel, Biorad II xi, 16 x 16 cm gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 21 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Tissue lysate - other (Human plasma 1 in 4 dilution)
Loading amount
20 µg
Specification
Human plasma 1 in 4 dilution
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5 %

Abcam user community

Verified customer

Submitted Feb 21 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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