Recombinant
RabMAb

Recombinant Anti-Survivin antibody [EP2880Y] - BSA and Azide free (ab192675)

Overview

  • Product name

    Anti-Survivin antibody [EP2880Y] - BSA and Azide free
    See all Survivin primary antibodies
  • Description

    Rabbit monoclonal [EP2880Y] to Survivin - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IF, IP, Sandwich ELISAmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Survivin (N terminal). The exact sequence is proprietary.

  • Positive control

    • HeLa and Jurkat lysates; human urinary bladder carcinoma and human colonic adenocarcinoma tissues.
  • General notes

    Ab192675 is the carrier-free version of ab76424. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab192675 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 ab192675 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 Use at an assay dependent concentration. Detects a band of approximately 16 kDa (predicted molecular weight: 16 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Signal amplification system might be necessary or the use of polymerized HRP secondary antibodies.

Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Sandwich ELISA Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Goat polyclonal to Survivin (ab27468).

Can be paired for Sandwich ELISA with Goat polyclonal to Survivin (ab27468).

Target

  • Function

    Component of the chromosomal passenger complex (CPC), a complex that acts as a key regulator of mitosis. The CPC complex has essential functions at the centromere in ensuring correct chromosome alignment and segregation and is required for chromatin-induced microtubule stabilization and spindle assembly. The complex with RAN plays a role in mitotic spindle formation by serving as a physical scaffold to help deliver the RAN effector molecule TPX2 to microtubules. May play a role in neoplasia. May counteract a default induction of apoptosis in G2/M phase. Inhibitor of caspase-3 and caspase-7. Isoform 2 and isoform 3 do not appear to play vital roles in mitosis. Isoform 3 shows a marked reduction in its anti-apoptotic effects when compared with the displayed wild-type isoform.
  • Tissue specificity

    Expressed only in fetal kidney and liver, and to lesser extent, lung and brain. Abundantly expressed in adenocarcinoma (lung, pancreas, colon, breast, and prostate) and in high-grade lymphomas. Also expressed in various renal cell carcinoma cell lines.
  • Sequence similarities

    Belongs to the IAP family.
    Contains 1 BIR repeat.
  • Developmental stage

    Expression is cell cycle-dependent and peaks at mitosis.
  • Domain

    The BIR repeat is necessary and sufficient for HBXIP binding.
  • Post-translational
    modifications

    Ubiquitination is required for centrosomal targeting.
    In vitro phosphorylation at Thr-117 by AURKB/STK12 prevents interaction with INCENP and localization to mitotic chromosomes.
  • Cellular localization

    Cytoplasm. Nucleus. Chromosome. Chromosome > centromere. Cytoplasm > cytoskeleton > spindle. Localizes on chromosome arms and inner centromeres from prophase through metaphase and then transferring to the spindle midzone and midbody from anaphase through cytokinesis. Colocalizes with AURKB at mitotic chromosomes.
  • Information by UniProt
  • Database links

  • Alternative names

    • API4 antibody
    • Apoptosis inhibitor 4 antibody
    • Apoptosis inhibitor survivin antibody
    • Apoptosis inhibitor4 antibody
    • Baculoviral IAP repeat containing 5 antibody
    • Baculoviral IAP repeat containing protein 5 antibody
    • Baculoviral IAP repeat-containing protein 5 antibody
    • BIRC 5 antibody
    • BIRC5 antibody
    • BIRC5_HUMAN antibody
    • EPR 1 antibody
    • IAP4 antibody
    • Survivin variant 3 alpha antibody
    • SVV antibody
    • TIAP antibody
    see all

Images

  • ab76424 (purified) at 1:150 dilution (5µg) immunoprecipitating Survivin in Ramos whole cell lysate.
    Lane 1 (input): Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10µg
    Lane 2 (+): ab76424 & Ramos whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76424 in Ramos whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

  • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Survivin with purified ab76424 at 1:500 dilution (5 µg/ml) (red). Cells were fixed with 80% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human bladder carcinoma tissue sections labeling Survivin with Purified ab76424 at 1:1000 dilution (2.52 µg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using Tris/EDTA Buffer,PH9. Tissue was counterstained with Hematoxylin. ab97051 Goat Anti-Rabbit IgG H&L (HRP)
    secondary antibody was used at 1:500 dilution. PBS instead of the primary antibody was used as the negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

  • ab76424 staining Survivin 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

  • Immunofluorescent staining of MCF-7 cells labelling Bcl-2 with purified ab76424 at 1/1000. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with ab7291, a mouse anti-tubulin (1/1000) using ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary antibody. Nuclei counterstained with DAPI (blue). 

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

  • Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling Survivin with unpurified ab76424 at 1/100. Cells were fixed with 4% paraformaldehyde for 20 minutes at room temperature. Cells were incubated with the primary antibody overnight. An Alexa Fluor® 555-conjugated anti-rabbit IgG was used as the secondary antibody. Left - Survivin, Right - DAPI.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

  • Standard curve for Survivin (Analyte: ab87202); dilution range 1pg/ml to 1µg/ml using Capture Antibody ab27468 at 1µg/ml and Detector Antibody abRabbit monoclonal [EP2880Y] to Survivin (ab76424) at 0.5µg/ml. Concentration of Rabbit monoclonal [EP2880Y] to Survivin (ab76424) may vary from lot to lot; please use this curve as guideline.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

  • Unpurified ab76424 at 1/250 dilution staining Survivin in human urinary bladder carcinoma by Immunohistochemistry, Paraffin-embedded tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

  • Unpurified ab76424 at 1/250 dilution staining Survivin in human colonic adenocarcinoma by Immunohistochemistry, Paraffin-embedded tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

  • Overlay histogram showing HeLa cells stained with unpurified ab76424 (red line). The cells were fixed with 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 (ab76424, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HeLa cells fixed with 4% paraformaldehyde/permeabilized with 0.1% PBS-Tween 20 used under the same conditions.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

  • Unpurified ab76424 showing positive staining in breast carcinoma tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

  • Unpurified ab76424 showing positive staining in cervical carcinoma tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

  • Unpurified ab76424 showing positive staining in Fetal kidney tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

  • Unpurified ab76424 showing positive staining in fetal liver tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

  • Unpurified ab76424 showing positive staining in normal tonsil tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76424).

References

This product has been referenced in:

  • Chen S  et al. SOX2 regulates apoptosis through MAP4K4-Survivin signaling pathway in human lung cancer cells. Carcinogenesis 35:613-23 (2014). WB, IHC-P ; Mouse, Human . Read more (PubMed: 24233838) »
  • Selemetjev S  et al. Evaluation of survivin expression and its prognostic value in papillary thyroid carcinoma. Pathol Res Pract N/A:N/A (2013). Human . Read more (PubMed: 24199968) »
See all 4 Publications for this product

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