Anti-Survivin antibody [EP2880Y] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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).

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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).

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) (top) or PBMCs treated with 1μg/mL PHA for 24h followed by 20U/mL IL-2 for 48h (bottom), with ab76424 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were fixed and permeabilised with BD Cytofix/Cytoperm™ for 20 min. PBMCs were incubated for 30 min at 4°C in 1x PBS containing 10 μg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab76424 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 μl at 0.2 μg/ml (1/11100)) for 30 min at 4°C . The cells were simultaneously stained with CD45RO.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 4°C

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on CD4+ cells.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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).

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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).

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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).

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

Clone EP2880Y (ab192675) has been successfully conjugated by Abcam. This image was generated using Anti-Survivin antibody [EP2880Y] (Alexa Fluor® 647). Please refer to ab204464 for protocol details.

ab204464 staining Survivin in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab204464 at a 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at a 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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).

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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).

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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).

This image is courtesy of an abreview submitted by Kirk Mcmanus.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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 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).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

Clone EP2880Y (ab192675) has been successfully conjugated by Abcam. This image was generated using Anti-Survivin antibody [EP2880Y] (Alexa Fluor® 488). Please refer to ab204264 for protocol details.

ab204264 staining Survivin in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab204264 at a 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at a 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

Intracellular 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).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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).

• IP

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Immunoprecipitation - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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).

All lanes:

Immunoprecipitation - Anti-Survivin antibody [EP2880Y] (<a href='/en-us/products/primary-antibodies/survivin-antibody-ep2880y-ab76424'>ab76424</a>)

Predicted band size: 16 kDa

false

• WB

Lab

Western blot - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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

Blocking and diluting buffer and concentration : 5% NFDM /TBST

ab76424 has a low sensitivity issue in western blot. We recommend ab208938 as an alternative.

All lanes:

Western blot - Anti-Survivin antibody [EP2880Y] (<a href='/en-us/products/primary-antibodies/survivin-antibody-ep2880y-ab76424'>ab76424</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 16 kDa

false

Exposure time: 60s

• sELISA

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Sandwich ELISA - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

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).

• OI-RD Scanning

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OI-RD Scanning - Anti-Survivin antibody [EP2880Y] - BSA and Azide free (AB192675)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.