Anti-SF2 antibody [EPR8240]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SF2 antibody [EPR8240] (AB133689)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling SF2 with purified ab133689 at 1/50 dilution (8.3 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SF2 antibody [EPR8240] (AB133689)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue sections labeling SF2 with Purified ab133689 at 1/100 dilution (4.16 μg/ml). Heat mediated antigen retrieval was performed using Citrate buffer, pH 6.0. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SF2 antibody [EPR8240] (AB133689)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling SF2 with purified ab133689 at 1/40 dilution (10μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SF2 antibody [EPR8240] (AB133689)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling SF2 with unpurified ab133689 at 1/50 dilution.

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

• WB

Lab

Western blot - Anti-SF2 antibody [EPR8240] (AB133689)

All lanes:

Western blot - Anti-SF2 antibody [EPR8240] (ab133689) at 1/1000 dilution

Lane 1:

293T (Human embryonic kidney epithelial cell) whole cell lysates at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 3:

C6 (Rat glial tumor glial cell) whole cell lysates at 20 µg

Lane 4:

RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates at 20 µg

Lane 5:

K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates 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

Predicted band size: 28 kDa

Observed band size: 27 kDa

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• WB

Unknown

Western blot - Anti-SF2 antibody [EPR8240] (AB133689)

All lanes:

Western blot - Anti-SF2 antibody [EPR8240] (ab133689) at 1/1000 dilution

Lane 1:

293T cell lysate at 10 µg

Lane 2:

C6 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 28 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-SF2 antibody [EPR8240] (AB133689)

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.