Overview

  • Product name
  • Description
    Rabbit polyclonal to Cyclin B1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, Sandwich ELISA, IHC-P, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Cyclin B1 (C terminal).
    Database link: P14635

  • Positive control
    • Jurkat, Ls174T or HeLa cells. Breast carcinoma tissue section.

Properties

Applications

Our Abpromise guarantee covers the use of ab2949 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
ICC/IF Use at an assay dependent concentration.
Sandwich ELISA Use a concentration of 0.2 µg/ml. For sandwich ELISA, use this antibody as Detection at 0.2 µg/ml with Mouse monoclonal [7A9] to Cyclin B1 (ab49215) as Capture.
IHC-P 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP Use at 10 µg/mg of lysate.
WB Use at an assay dependent concentration. Detects a band of approximately 60 kDa (predicted molecular weight: 48 kDa).

Target

Images

  • Standard Curve for Cyclin B1 (Analyte: Cyclin B1 protein (His tag) (ab40591)); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal [7A9] to Cyclin B1 (ab49215) at 0.2µg/ml and Detector Antibody Rabbit polyclonal to Cyclin B1 (ab2949) at 0.2µg/ml.
  • Breast carcinoma stained with ab2949
  • ab2949 staining Cyclin B1 in mouse MEF cells by Immunocytochemistry/ immunoflurescence. Cells were fixed with paraformaldehyde and blocking with 2% serum was performed for 2 hours at 250C. Samples were incubated with primary antibody (1/100: in 1% BSA, 0.3% Triton X-100 in PBS) for 18 hours at 4°C. An Alexa Fluor®594-conjugated goat polyclonal to rabbit IgG was used diluted 1/1000 as secondary antibody. Red is cyclinB1 staining, blue is DAPI staining DNA for nuclei, green is vinculin staining.

    See Abreview

References

This product has been referenced in:
  • Burger K & Gullerova M Nuclear re-localization of Dicer in primary mouse embryonic fibroblast nuclei following DNA damage. PLoS Genet 14:e1007151 (2018). Read more (PubMed: 29394246) »
  • Meng X  et al. MicroRNA-125b-1-3p mediates intervertebral disc degeneration in rats by targeting teashirt zinc finger homeobox 3. Exp Ther Med 15:2627-2633 (2018). Read more (PubMed: 29456666) »
See all 10 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (Human Cell lysate - whole cell (Epithelial OVCAR c)
Gel Running Conditions
Reduced Denaturing (10 % SDS-PAGE)
Loading amount
100 µg
Treatment
Perifosine 0, 12, 24 hr
Specification
Human Cell lysate - whole cell (Epithelial OVCAR c
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Apr 17 2017

Answer

Thank you for contacting us.

Unfortunately the immunogen sequence is confidential. It is located within aa 370-433 of P14635 (Uniprot).

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for confirming that.

I have generated the testing discount code as follows:

Discount code: xxxxxx

Expiration date: 5th December 2012

The terms and conditions of the testing discount scheme can be found from the following link:

https://www.abcam.com/collaboratordiscount

I hope this information has been of help. If you have any further questions, please do not hesitate to ask.

Read More

Answer

Thank you for contacting us and sorry for the delay in getting back to you.

As requested, I have looked into the sequence homology between the immunogen used to raise antibodies mentioned and the relevant canineprotein sequence. I have to mention that all the caninesequences I have found havenot been fully characterised but predicted in SwissProt, I cannot therefore be certain of the homology predictions. This is what I have found:

Anti-Cyclin B1 antibody (ab2949): immunogen used taken from C-terminal sequence of human cyclin B1. This shares 100% homology with the canine sequence (SwissProt reference E2R3S2).

Anti-PLK1 antibody [Mixed clones, 3G251] (ab14210): immunogen used fusion protein of residues 402-603 of human PLK1. This shares 97% homology with the canine protein (SwissProt reference E2RNE0)

Anti-PLK1 antibody [36-298] (ab17057): immunogen used His-PLK1 full length purified from Sf9 cells. The epitope of the antibody lies between the residues 330-370. This region shares 85% homologywith the canine protein (SwissProt reference E2RNE0)

All three antibodies would therefore be predicted to react with the canine protein. However, as we have not tested this ourselves and have had no report of other customers doing so either, we would not be able to guarantee this.

I hope this information has been of help. If you have any further questions, please do not hesitate to contact us again.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (Liver)
Loading amount
50 µg
Specification
Liver
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Sep 05 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver)
Loading amount
50 µg
Specification
Liver
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Aug 24 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (MEF)
Specification
MEF
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 28 2009

Answer

Thank you for your e-mail and forwarding more information about your Western blot assay. We are very sorry to hear that you are having problem with this antibody. We have searched our database and found that this is a popular selling product and your feedback is the first we have received about it not working. Therefore, at this stage, we would suggest that there is either a problem with the vial you received, or modifications to your protocol are needed to obtain a positive result. 1. Make sure that you prepare the samples with a lysis buffer containing proteinase inhibitors. Usually, lower bands indicate that there may be degraded protein in the samples. 2. You mentioned in your e-mail that you loaded 50/100 mg protein per lane. This is too much. We recommend using 25-30 microgram/ml (ug/ml). 3. We would like to know the band size what you have. Since there is no label on the MW bands on the gel, it is very difficult to assess the band size. 4. Have you tested the protein transfer? Have you run a secondary antibody 'only' experiment to see if the non-specific bands are due to the secondary antibody? We hope this will help.

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

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