Name: Anti-Cyclin E2 antibody [E142]
Brand: RabMAb
SKU: ab32103
Rating: 4 (5 reviews)

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Western blot - Anti-Cyclin E2 antibody [E142] (ab32103)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [E142] to Cyclin E2
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
Reacts with: Human

Product name

Anti-Cyclin E2 antibody [E142]
See all Cyclin E2 primary antibodies
Description

Rabbit monoclonal [E142] to Cyclin E2
Host species

Rabbit
Specificity

ab32103 recognises Cyclin E2.
Tested applications

Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
Unsuitable for: IP
Species reactivity

Reacts with: Human
Predicted to work with: Mouse
Immunogen

Synthetic peptide within Human Cyclin E2 aa 350 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: O96020
Positive control
- Human breast carcinoma, HeLa cells lysates, Jurkat whole cell lysate (ab7899)
General notes
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

IgG fraction
Clonality

Monoclonal
Clone number

E142
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- HeLa whole cell lysate (ab29545)

Our Abpromise guarantee covers the use of ab32103 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		1/500. Predicted molecular weight: 45 kDa.
IHC-P		Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF		1/2000. For unpurified use at 1/100 - 1/250.
Flow Cyt		1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Application notes

Is unsuitable for IP.

Relevance

The human Cyclin E2 gene encodes a 404 amino acid protein that is most closely related to Cyclin E. Cyclin E2 mRNA levels peaks at the G1 / S transition. Cyclin E2 associates with Cdk2 in a functional kinase complex that is inhibited by both p27 (Kip1) and p21 (Cip1). Cyclin E2 / Cdk2 phosphorylates histone H1 in vitro. G1 cyclin E controls the initiation of DNA synthesis by activating CDK2. Abnormally high levels of cyclin E expression have frequently been observed in human cancers. Unlike Cyclin E1, which is expressed in great majority of proliferating normal and neoplastically transformed cells, Cyclin E2 levels are low to undetectable in non transformed cells and increase significantly in neoplasm derived cells.
Cellular localization

Nuclear
Database links
- Entrez Gene: 9134 Human
- Entrez Gene: 12448 Mouse
- Omim: 603775 Human
- SwissProt: O96020 Human
- SwissProt: Q7TMS8 Mouse
- SwissProt: Q9Z238 Mouse
- Unigene: 567387 Human
Alternative names
- CCN E2 antibody
- CCNE 2 antibody
- CCNE2 antibody
- CCNE2 protein antibody
- CYC E2 antibody
- CYCE 2 antibody
- CYCE2 antibody
- CyclinE2 antibody
- G1/S specific cyclin E2 antibody
see all

Western blot - Anti-Cyclin E2 antibody [E142] (ab32103)

Anti-Cyclin E2 antibody [E142] (ab32103) at 1/500 dilution + HeLa cell lysate

Predicted band size: 45 kDa
Observed band size: 50 kDa
why is the actual band size different from the predicted?
Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E2 antibody [E142] (ab32103)

Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling Cyclin E2 with purified ab32103 at 1/2000. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Nuclei counterstained with DAPI (blue).

Secondary Only Control: PBS was used instead of the primary antibody as the negative control.
Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E2 antibody [E142] (ab32103)

Immunofluorescent analysis of Cyclin E2 expression in HeLa cell culture using 1/100 ab32103.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclin E2 antibody [E142] (ab32103)

Immunohistochemical analysis of Cyclin E2 expression in paraffin embedded human breast carcinoma using 1/50 ab32103.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cytometry - Anti-Cyclin E2 antibody [E142] (ab32103)

Overlay histogram showing HeLa cells stained with ab32103 (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 (ab32103, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
Immunocytochemistry/ Immunofluorescence - Anti-Cyclin E2 antibody [E142] (ab32103)This image is courtesy of an abreview by Kirk Mcmanus

ab32103 staining Cyclin E2 in the Hela cell line from Human cervix by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. Ab150081 (1/200) was used as the secondary antibody.
See Abreview
Anti-Cyclin E2 antibody [E142] (ab32103)

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.

Click here to view the general protocols

Publishing research using ab32103? Please let us know so that we can cite the reference in this datasheet.

ab32103 has been referenced in 14 publications.

Zhao F et al. MicroRNA-133b suppresses bladder cancer malignancy by targeting TAGLN2-mediated cell cycle. J Cell Physiol 234:4910-4923 (2019). PubMed: 30317571
Liu J et al. Downregulation of LncRNA-XIST inhibited development of non-small cell lung cancer by activating miR-335/SOD2/ROS signal pathway mediated pyroptotic cell death. Aging (Albany NY) 11:7830-7846 (2019). PubMed: 31553952
Hou Y et al. SKA3 Promotes tumor growth by regulating CDK2/P53 phosphorylation in hepatocellular carcinoma. Cell Death Dis 10:929 (2019). PubMed: 31804459
Ganguly S et al. Pathobiology of cigarette smoke-induced invasive cancer of the renal pelvis and its prevention by vitamin C. Toxicol Rep 5:1002-1010 (2018). PubMed: 30338226
Lv W et al. KIAA0101 inhibition suppresses cell proliferation and cell cycle progression by promoting the interaction between p53 and Sp1 in breast cancer. Biochem Biophys Res Commun 503:600-606 (2018). PubMed: 29902451

View all Publications for this product

Customer reviews and Q&As

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1-8 of 8 Abreviews or Q&A

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (HeLa)

Permeabilization

Yes - 0.5% Triton X100 in PBS

Specification

HeLa

Fixative

Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Aug 25 2017

Question

IHC-P and chromosome spreads
received warm and tried it, but no signal
worked with same Ab before and it was fine
mouse prostate tissue
AR: Tris pH 9
Ab: 1/100 o/n
block: donkey serum
2nd: OK

Abcam community

Verified customer

Asked on Aug 07 2012

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.
I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx. The estimated delivery date is xxx.
To check the status of the order please contact our Customer Service team and reference this number.
Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
I wish you the best of luck with your research.

Abcam Scientific Support

Answered on Aug 07 2012

Application

Western blot

Sample

Rat Tissue lysate - whole (Liver)

Loading amount

50 µg

Specification

Liver

Gel Running Conditions

Reduced Denaturing (12%)

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 28 2011

Application

Western blot

Sample

Mouse Cell lysate - whole cell (Hepa 1-6)

Loading amount

50 µg

Specification

Hepa 1-6

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 23 2011

Application

Western blot

Sample

Human Cell lysate - whole cell (HepG2)

Loading amount

50 µg

Specification

HepG2

Gel Running Conditions

Reduced Denaturing (12)

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Mrs. Sadaf Sultan

Verified customer

Submitted Aug 11 2011

Application

Western blot

Sample

Human Cell lysate - whole cell (HeLa cell line)

Loading amount

50 µg

Specification

HeLa cell line

Gel Running Conditions

Reduced Denaturing

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Dr. Shakil Ahmad

Verified customer

Submitted Aug 11 2011

Question

BATCH NUMBER 319539 ORDER NUMBER PO-00843 DESCRIPTION OF THE PROBLEM Cytoplasmic staing and heavy background SAMPLE Breast carcinoma PRIMARY ANTIBODY 1:50 for 2hours DETECTION METHOD Streptavidin HRP label POSITIVE AND NEGATIVE CONTROLS USED Pos-Human Breast carcinoma Neg- Normal rabbit IgG Fraction ANTIBODY STORAGE CONDITIONS 4 degrees FIXATION OF SAMPLE Archived formalin fixed paraffin embedded ANTIGEN RETRIEVAL Pressure cooker and pH 6.0 modified citrate PERMEABILIZATION STEP none BLOCKING CONDITIONS Avidin/Biotin blocks SECONDARY ANTIBODY Biotinylated link HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5+ HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? No WHAT STEPS HAVE YOU ALTERED? I have taken over this project from someone else. I have no other information regarding staining parameters ADDITIONAL NOTES I am looking to steer away from the avidin/biotin system and am going to try a pH 9.0 Tris-HCL retrieval in gentle waterbath 20mins and incubate overnight at 4 degrees. There is too much cytoplasmic staining and some stromal background I must get rid of. I just wanted an idea of Abcam’s staining method for IHC so I don’t waste anymore precious tissue or reagent. And time!

Abcam community

Verified customer

Asked on Jul 09 2007

Answer

Thank you for your enquiry. I can confirm that this product does not require any specific protocols and the general IHC-P and antigen retrieval protocols found on our website (can also be accessed on the datasheet) will be able to help you. From the protocols you provided, I agree that you should try incubating at 4oC overnight to ensure proper and sufficient binding to the protein of interest. You can also try increasing the primary antibody dilution to 1:100 ~ 1:1000. The recommended dilution we have on the datasheet is just a guideline and optimal dilutions/concentrations should be determined by the end user. Can I also suggest running a no primary experiment? Sometimes, the problem lies with the secondary antibody and could be binding non-specifically to your sample. I hope the information above will helpful. Please do not hesitate to contact me again if you face problems when using this product.

Abcam Scientific Support

Answered on Jul 09 2007

Question

I have a customer enquiring about product# ab32103, and want to know the concentration of this antibody. Could you please advise the estimated concentration for this product?

Abcam community

Verified customer

Asked on Aug 28 2006

Answer

Thank you for your enquiry. The approximate concentration of this antibody (ab32103) is 90ug/ml. I hope this is helpful. Please don't hesitate to contact us again if you require more information.

Abcam Scientific Support

Answered on Aug 30 2006

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Positive Controls

Applications

Target

Relevance

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

References (14)

Customer reviews and Q&As

Immunocytochemistry/ Immunofluorescence abreview for Anti-Cyclin E2 antibody [E142]

Western blot abreview for Anti-Cyclin E2 antibody [E142]

Western blot abreview for Anti-Cyclin E2 antibody [E142]

Western blot abreview for Anti-Cyclin E2 antibody [E142]

Western blot abreview for Anti-Cyclin E2 antibody [E142]