Anti-Estrogen Inducible Protein pS2 antibody [EPR3972]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Inducible Protein pS2 antibody [EPR3972] (AB92377)

ab92377 staining Estrogen Inducible Protein pS2 in Formalin-fixed, Paraffin-embedded Human breast carcinoma tissue (A) and Human stomach tissue (B) at 1/100 dilution. Detection : DAB staining.

Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Inducible Protein pS2 antibody [EPR3972] (AB92377)

ab92377 showing positive staining in Ductal infiltrating breast carcinoma tissue.

Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Inducible Protein pS2 antibody [EPR3972] (AB92377)

ICC/IF image of ab92377 stained MCF7 cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab92377, 1/100 dilution) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Estrogen Inducible Protein pS2 antibody [EPR3972] (AB92377)

Intracellular Flow Cytometry analysis of MCF-7 (human breast carcinoma) cells labeling Estrogen Inducible Protein pS2 with unpurified ab92377 at 1/20 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Inducible Protein pS2 antibody [EPR3972] (AB92377)

ab92377 showing negative staining in Normal breast tissue.

Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Inducible Protein pS2 antibody [EPR3972] (AB92377)

ab92377 showing positive staining in Ovarian carcinoma tissue.

Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-Estrogen Inducible Protein pS2 antibody [EPR3972] (AB92377)

Estrogen Inducible Protein pS2 was immunoprecipitated from 0.35 mg MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10 μg with ab92377 at 1/50 dilution (2μg). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10 μg

Lane 2 : ab92377 IP in MCF7 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab92377 in MCF7 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Estrogen Inducible Protein pS2 antibody [EPR3972] (ab92377)

Predicted band size: 9 kDa

Observed band size: 9 kDa

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• WB

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Western blot - Anti-Estrogen Inducible Protein pS2 antibody [EPR3972] (AB92377)

All lanes:

Western blot - Anti-Estrogen Inducible Protein pS2 antibody [EPR3972] (ab92377) at 1/1000 dilution

Lane 1:

Human stomach lysate at 10 µg

Lane 2:

MCF-7 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 18 kDa,9 kDa

Observed band size: 22 kDa

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• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Inducible Protein pS2 antibody [EPR3972] (AB92377)

ab92377 showing negative staining in Normal kidney tissue.

Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

• OI-RD Scanning

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OI-RD Scanning - Anti-Estrogen Inducible Protein pS2 antibody [EPR3972] (AB92377)

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.