Overview

  • Product name
  • Description
    Rabbit polyclonal to Centrin 1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Dog, Human
    Predicted to work with: Xenopus laevis, Drosophila melanogaster
  • Immunogen

    Synthetic peptide conjugated to KLH, corresponding to amino acids 157-172 of Human Centrin 1, with N-terminally added lysine. This sequence is highly conserved in centrin 1 of many species, e.g. mouse, drosophila, and Xenopus. It is also found in human and mouse centrin 2, but is not found in centrin 3.

  • Positive control
    • HeLa cell line.
  • General notes
    Storage in frost-free freezers is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use.

Properties

Applications

Our Abpromise guarantee covers the use of ab11257 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
WB 1/2000. Predicted molecular weight: 21 kDa. 1/2000, this concentration is recommended using a nuclear extract of the human epitheloid carcinoma HeLa cell line. Predicted molecular weight: 21 kDa.
ICC/IF 1/200. PubMed: 17299043

Suggested working dilution of 1/200 (see PMID: 17299043).

Methanol fixation may be preferable for this antibody based on publications and customer feedback.

Target

  • Function
    Plays a fundamental role in microtubule-organizing center structure and function.
  • Sequence similarities
    Belongs to the centrin family.
    Contains 4 EF-hand domains.
  • Cellular localization
    Cytoplasm > cytoskeleton > centrosome. Centrosome of interphase and mitotic cells.
  • Information by UniProt
  • Database links
  • Alternative names
    • Caltractin antibody
    • Caltractin isoform 2 antibody
    • CEN 1 antibody
    • CEN1 antibody
    • Centrin EF hand protein 1 antibody
    • Centrin-1 antibody
    • Centrin1 antibody
    • CETN 1 antibody
    • CETN antibody
    • CETN1 antibody
    • CETN1_HUMAN antibody
    • EF hand protein 1 antibody
    see all

Images

  • ab11257 staining Centrin 1 in mouse embryonic fibroblast cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with Ethanol (-200C) and blocking with 3% BSA/PBS  was done for 30 minutes at 220C. Samples were incubated with primary antibody (1/500) in in 3% BSA /0.1 % saponin/PBS for 1 hour at RT. An undiluted Alexa Fluor®546-conjugated donkey polyclonal to goat IgG (H&L) was used at dilution at 1/500 as secondary antibody. In the image, Green staining represents gamma tubulin, red - centrin.

    See Abreview

References

This product has been referenced in:
See all 24 Publications for this product

Customer reviews and Q&As

1-10 of 17 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HCT116)
Permeabilization
Yes - 0.5% Triton-X100 in PBS
Specification
HCT116
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Jan 16 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Xenopus laevis Cell (gastrocoel roof plate)
Permeabilization
Yes - 0.1% Triton X-100
Specification
gastrocoel roof plate
Blocking step
3% BSA + 3% serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 29 2015

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 23°C
Sample
Mouse Cell (Liver)
Specification
Liver
Permeabilization
Yes - 0.5% Triton in PBS
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 13 2014

Answer

Thank you for your response.

I am not sure what IB represents but assume the customer would mean either IP (Immunoprecipitation) or WB (Western blot) application? Is that correct?

I regret to inform you that I am not convinced that this antibody (ab99337) is faulty. However, I appreciate your long-term relationship with this customer and try to help you to close this case soon.

Here are a few comments I would like to make and share:

Multiple bands could be due to several factors (blocking, protein degradation, secondary antibody). Since the non-specific bands are lower range than the expected band size for CP110, it may well be that the bands are degradation product.

Unfortunately, I have limited information about the experiments but in future in similar complaint the following details would be advised to provide in order to get sufficient technical support

What Lane 1 and 2 represent? Identical samples? Or different preparation?
How was the blocking applied (blocking agent, time and temperature)?
How much protein was loaded?
Has "No primary" (only secondary) antibody control was used to rule out any non-specific signal due to the secondary antibody?



Please be advised that there are two isoforms (O43303-1 and O43303-2) which have been described in the literature and it would be advisable to check which one the customer’s samples express. Your customer could find some further information about this target at this site:

http://www.uniprot.org/uniprot/O43303

Furthermore, CCP110 has different subunits and the CCP110 protein can interact with other proteins such CALM1, CETN2, CEP76, CEP97, NEURL4 and CCNF. For this reason, it is essential to know if the experiments were carried out under naturing or denaturing/and reducing conditions. This information is also missing in the last e-mail and would be very important to know.

Currently, we have two alternative antibodies against CP110 ab99338 and ab91415 and can offer one free of charge replacement vial. Could you please discuss the content of this e-mail with your customer and you let me know which product he wishes to test.

Thank you for your continued cooperation.

Read More

Answer

Thank you for your response.

Unfortunately, the two image files can't be opened so I am unable to take a look at the signal.

Tested and Guaranted Applications:

ab11257 - WB, ICC/IF

ab99337 - WB and IP - this has not been tested in immunostaining.

Since ab99337 has not been tested in immunostaining, I am very sorry but our Abpromise does not apply for this antibody.

Regarding ab11257, the protocol looks fine though it would be interesting to know if the cultured HeLa cells are from the exponential phase of growth or from a confluent culture. No information is provided. Furthermore, blocking non-specific bindings with 10% BSA may be a bit too strong with cultured cells.

I can offer a new free of charge vial of 11257 but not for the other product ab99337 which has not been tested for ICC/IF yet.

Could you please discuss our policy with the customer to make sure he understands for which product we can offer the Abpromise.

I look forward to hearing from you soon.

Read More

Answer

Thank you for your enquiry regarding our two products. I am very sorry to hear that you are having problems with this antibody.

As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

Tested and Guaranted Applications:

ab11257 - WB, ICC/IF

ab99337 - WB and IP - this has not been tested in immunostaining

Could you please provide some details of the protocol used and complete the following form (attached as a word document). It would be much appreciated if you could attach an image to the response.

- Batch/Order numbers,

- Species and type of the sample,

- Cells or tissue section used,

- Fixation method

Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

Read More

Answer

Thank you very much for your recent review of ab11257. Your review is now published on our website.
Since the results were below average, I'm contacting you to see if we can help resolve this problem. I would recommend reducing the amount of protein loaded to 20-30 ug, and you can also block the membrane overnight at 4C then incubate with the primary antibody for 1 hour at room temperature. These steps may help reduce the non-specific bands on the blot.
We do fully guarantee this antibody to work with human samples in Western blot, so if you would like a replacement antibody or a credit or refund, please let me know your original order number or PO number and I'll be happy to set that up for you.
I look forward to hearing from you. If there is anything else that we can do for you, please let me know and I'll be happy to help.

Read More
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (Hela)
Loading amount
50 µg
Specification
Hela
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Aug 03 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Hela)
Specification
Hela
Fixative
Methanol
Permeabilization
No
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Aug 03 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Embryonic stem)
Specification
Embryonic stem
Fixative
Methanol
Permeabilization
Yes - 0.1% triton-X-100
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 25°C

Prof. Reiter Lab

Verified customer

Submitted Apr 19 2010

1-10 of 17 Abreviews or Q&A

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

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