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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)

Reviews (1) Submit a question References (1)
Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
  • Flow Cytometry (Intracellular) - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
  • Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
  • Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
  • Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
  • Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
  • Immunocytochemistry/ Immunofluorescence - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
  • Immunocytochemistry/ Immunofluorescence - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
  • Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17865] to SA2 - C-terminal
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-SA2 antibody [EPR17865] - C-terminal
    See all SA2 primary antibodies

  • Description

    Rabbit monoclonal [EPR17865] to SA2 - C-terminal

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, HCT116, MCF-7, K562, C6, Raw264.7 and NIH3T3 cell lysates; Human fetal brain and Mouse spleen lysates; IHC-P: Human breast carcinoma, human tonsil, mouse and rat spleen tissue; IF: MCF-7 and K562 cells.

  • General notes

    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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab201451 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 (Intra)
1/25000.
WB (1)
1/1000. Detects a band of approximately 141 kDa (predicted molecular weight: 141 kDa).
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/500.
Notes
Flow Cyt (Intra)
1/25000.
WB
1/1000. Detects a band of approximately 141 kDa (predicted molecular weight: 141 kDa).
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/500.

Target

  • Function

    Component of cohesin complex, a complex required for the cohesion of sister chromatids after DNA replication. The cohesin complex apparently forms a large proteinaceous ring within which sister chromatids can be trapped. At anaphase, the complex is cleaved and dissociates from chromatin, allowing sister chromatids to segregate. The cohesin complex may also play a role in spindle pole assembly during mitosis.

  • Sequence similarities

    Belongs to the SCC3 family.
    Contains 1 SCD (stromalin conservative) domain.

  • Post-translational
    modifications

    Phosphorylated by PLK. The large dissociation of cohesin from chromosome arms during prophase is partly due to its phosphorylation.

  • Cellular localization

    Nucleus. Chromosome. Chromosome > centromere. Associates with chromatin. Before prophase it is scattered along chromosome arms. During prophase, most of cohesin complexes dissociate from chromatin probably because of phosphorylation by PLK, except at centromeres, where cohesin complexes remain. At anaphase, the RAD21 subunit of cohesin is cleaved, leading to the dissociation of the complex from chromosomes, allowing chromosome separation. In germ cells, cohesin complex dissociates from chromatin at prophase I, and may be replaced by a meiosis-specific cohesin complex.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • bA517O1.1 antibody
    • Cohesin Subunit SA 2 antibody
    • Cohesin subunit SA-2 antibody
    • DKFZp686P168 antibody
    • DKFZp781H1753 antibody
    • FLJ25871 antibody
    • SA 2 antibody
    • SA-2 antibody
    • SA2 antibody
    • SCC3 homolog 2 antibody
    • SCC3B antibody
    • STAG 2 antibody
    • stag2 antibody
    • STAG2_HUMAN antibody
    • Stromal antigen 2 antibody
    see all

Images

  • Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    All lanes : Anti-SA2 antibody [EPR17865] - C-terminal (ab201451) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : STAG2 knockout HeLa cell lysate
    Lane 3 : HCT116 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 141 kDa
    Observed band size: 141 kDa



    Lanes 1-3: Merged signal (red and green). Green - ab201451 observed at 141 kDa. Red - loading control ab8245 observed at 36 kDa.

    ab201451 Anti-SA2 antibody [EPR17865] - C-terminal was shown to specifically react with SA2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265461 (knockout cell lysate ab257707) was used. Wild-type and SA2 knockout samples were subjected to SDS-PAGE. ab201451 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

  • Flow Cytometry (Intracellular) - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    Flow Cytometry (Intracellular) - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)

    Intracellular Flow Cytometry analysis of K562 (human chronic myelogenous leukemia) labelling SA2 with purified ab201451 at 1/25000 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Alexa Fluor® 488 goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies. 

     

     

     

  • Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    All lanes : Anti-SA2 antibody [EPR17865] - C-terminal (ab201451) at 1/10000 dilution

    Lane 1 : MCF-7 (Human breast adenocarcinoma cell line) cell lysate
    Lane 2 : K562 (Human chronic myelogenous leukemia cells from bone marrow) 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: 141 kDa
    Observed band size: 141 kDa


    Exposure time: 3 minutes


    5% NFDM/TBST: Blocking and diluting buffer.

  • Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    Anti-SA2 antibody [EPR17865] - C-terminal (ab201451) at 1/1000 dilution + Human fetal brain lysate at 10 µg

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

    Predicted band size: 141 kDa
    Observed band size: 141 kDa


    Exposure time: 1 minute


    5% NFDM/TBST: Blocking and diluting buffer.

  • Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    All lanes : Anti-SA2 antibody [EPR17865] - C-terminal (ab201451) at 1/10000 dilution

    Lane 1 : C6 (Rat glial tumor cells) cell lysate
    Lane 2 : Raw264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) cell lysate
    Lane 3 : NIH 3T3 (Mouse embyro fibroblast cells) 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: 141 kDa
    Observed band size: 141 kDa


    Exposure time: 3 minutes


    5% NFDM/TBST: Blocking and diluting buffer.

  • Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    Western blot - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    Anti-SA2 antibody [EPR17865] - C-terminal (ab201451) at 1/1000 dilution + mouse spleen lysate at 10 µg

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

    Predicted band size: 141 kDa
    Observed band size: 141 kDa


    Exposure time: 1 minute


    5% NFDM/TBST: Blocking and diluting buffer.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)

    Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling SA2 using ab201451 at 1/2000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counterstain: Hematoxylin.
    Inset image: negative control obtained using PBS instead of ab201451, and secondary antibody only.
    Note: Nuclear staining on Human breast carcinoma tissue was observed.

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)

    Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling SA2 using ab201451 at 1/2000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counterstain: Hematoxylin.
    Inset image: negative control obtained using PBS instead of ab201451, and secondary antibody.
    Note: Nuclear staining on Human tonsil tissue was observed.

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)

    Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling SA2 using ab201451 at 1/2000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counterstain: Hematoxylin.
    Inset image: negative control obtained using PBS instead of ab201451, and secondary antibody.
    Note: Nuclear staining on mouse spleen tissue was observed.

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)

    Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling SA2 using ab201451 at 1/2000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counterstain: Hematoxylin.
    Inset image: negative control obtained using PBS instead of ab201451, and secondary antibody.
    Note: Nuclear staining on rat spleen tissue was observed.

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

  • Immunocytochemistry/ Immunofluorescence - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    Immunocytochemistry/ Immunofluorescence - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling SA2 with ab201451 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing nuclear staining on MCF7 cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:
    1. ab201451 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    Immunocytochemistry/ Immunofluorescence - Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling SA2 with ab201451 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing nuclear staining on K562 cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:
    1. ab201451 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)
    Anti-SA2 antibody [EPR17865] - C-terminal (ab201451)

References (1)

Publishing research using ab201451? Please let us know so that we can cite the reference in this datasheet.

ab201451 has been referenced in 1 publication.

  • Lovejoy CA  et al. ATRX affects the repair of telomeric DSBs by promoting cohesion and a DAXX-dependent activity. PLoS Biol 18:e3000594 (2020). PubMed: 31895940

Customer reviews and Q&As

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Western blot abreview for Anti-SA2 antibody [EPR17865] - C-terminal

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (ML-2 cells)
Gel Running Conditions
Reduced Denaturing (4-20% tris-glycin gel)
Loading amount
40 µg
Specification
ML-2 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted Oct 01 2020

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