Recombinant
RabMAb

Recombinant Anti-Cyclin A2 antibody [EPR17351] (ab181591)

Rabbit recombinant monoclonal Cyclin A2 antibody [EPR17351]. Validated in WB, IHC, ICC/IF and tested in Mouse, Rat, Human. Cited in 12 publication(s). Independently reviewed in 1 review(s).

Overview

  • Product name

    Anti-Cyclin A2 antibody [EPR17351]
    See all Cyclin A2 primary antibodies
  • Description

    Rabbit monoclonal [EPR17351] to Cyclin A2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Mouse Cyclin A2 aa 1-200. The exact sequence is proprietary.
    Database link: P51943

  • Positive control

    • WB: Jurkat, C6, RAW 264.7 and PC-12 whole cell lysates; HeLa Untreated, asynchronous cells, HeLa G1/S arrested cells (thymidine treatment) and HeLa G2/M arrested cells (sequential thymidine and nocodazole treatments) cell lysates; Human tonsil and fetal kidney lysates; Rat spleen lysate. IHC-P: Human tonsil, Human cervix carcinoma, rat colon and mouse endometrium tissues. ICC/IF: HeLa cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab181591 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 1/500.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/2000. Detects a band of approximately 50 kDa (predicted molecular weight: 47 kDa).

Target

Images

  • Anti-Cyclin A2 antibody [EPR17351] (ab181591) at 1/20000 dilution + Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 47 kDa
    Observed band size: 50 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Cyclin A2 with ab181591 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear and weakly cytoplasmic staining on HeLa cells. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [EPR17351]- Loading Control (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab181591 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/500 dilution.
    -ve control 2: Anti-alpha Tubulin antibody [EPR17351]- Loading Control (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Cyclin A2 with ab181591 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and weak cytoplasmic staining on germinal center cells of Human tonsil tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • All lanes : Anti-Cyclin A2 antibody [EPR17351] (ab181591) at 1/20000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) lysate; Untreated, asynchronous cells (ab136811)
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) lysate; G1/S arrested cells (thymidine treatment) (ab136811)
    Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) lysate; G2/M arrested cells (sequential thymidine and nocodazole treatments) (ab136811)

    Lysates/proteins at 1 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated or Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 47 kDa
    Observed band size: 50 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Cyclin A2 is down regulated at the G2/M phase.

  • Anti-Cyclin A2 antibody [EPR17351] (ab181591) + Human tonsil lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 47 kDa
    Observed band size: 50 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-Cyclin A2 antibody [EPR17351] (ab181591) at 1/2000 dilution + Human fetal kidney lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 47 kDa
    Observed band size: 50 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The smaller band is due to a tissue-specific splice variant, PMID 22745723.

  • All lanes : Anti-Cyclin A2 antibody [EPR17351] (ab181591) at 1/2000 dilution

    Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 47 kDa
    Observed band size: 50 kDa why is the actual band size different from the predicted?


    Exposure time: 3 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-Cyclin A2 antibody [EPR17351] (ab181591) at 1/2000 dilution + Rat spleen lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 47 kDa
    Observed band size: 50 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The smaller band is due to a tissue-specific splice variant, PMID 22745723.

  • Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling Cyclin A2 with ab181591 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and weak cytoplasmic staining on some tumor cells in Human cervix carcinoma tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Cyclin A2 with ab181591 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and weak cytoplasmic staining on a proportion of epithelial cells in rat colon tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded mouse endometrium tissue labeling Cyclin A2 with ab181591 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and weak cytoplasmic staining on a proportion of epithelial cells in mouse endometrial tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

References

This product has been referenced in:

  • Ren S  et al. Microvesicles from human adipose stem cells promote wound healing by optimizing cellular functions via AKT and ERK signaling pathways. Stem Cell Res Ther 10:47 (2019). Read more (PubMed: 30704535) »
  • Ji W  et al. Combined Androgen receptor blockade overcomes the resistance of breast cancer cells to palbociclib. Int J Biol Sci 15:522-532 (2019). Read more (PubMed: 30745839) »
See all 22 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Derived Breast Cancer Cell Lines)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
30 µg
Specification
Mouse Derived Breast Cancer Cell Lines
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C

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Submitted Jul 07 2017

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