Anti-SUN2 antibody [EPR6557]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SUN2 antibody [EPR6557] (AB124916)

ab124916 staining SUN2 in Human colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/500). An HRP-conjugated Goat anti-rabbit IgG, ab97051 (1/500), was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SUN2 antibody [EPR6557] (AB124916)

Immunofluorescence staining of SUN2 using ab124916 in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton-X-100 (in PBS) for 5 mins 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 ab124916 at 0.2 μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SUN2 antibody [EPR6557] (AB124916)

ab124916, unpurified, at a 1/250 dilution, staining SUN2 in paraffin embedded Human ovarian tissue by Immunohistochemistry.

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

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SUN2 antibody [EPR6557] (AB124916)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling SUN2 (red) with ab124916 at a 1/30 dilution. Cells were fixed with 80% methanol and permeabilized with 0.1% Tween-20. A goat anti-rabbit IgG (Alexa Fluorr^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SUN2 antibody [EPR6557] (AB124916)

ab124916, unpurified, at a 1/250 dilution, staining SUN2 in paraffin embedded Human lung tissue by Immunohistochemistry.

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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SUN2 antibody [EPR6557] (AB124916)

Immunofluorescence staining of SUN2 using ab124916 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton-X-100 (in PBS) for 5 mins 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 ab124916 at 1.0 μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SUN2 antibody [EPR6557] (AB124916)

ab124916 staining SUN2 in mouse testis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/500). An HRP-conjugated goat anti-rabbit IgG, ab97051 (1/500) was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.

• WB

Unknown

Western blot - Anti-SUN2 antibody [EPR6557] (AB124916)

All lanes:

Western blot - Anti-SUN2 antibody [EPR6557] (ab124916) at 1/1000 dilution

Lane 1:

Human fetal muscle lysate at 10 µg

Lane 2:

Saos-2 lysate at 10 µg

Lane 3:

HeLa lysate at 10 µg

Lane 4:

Jurkat lysate at 10 µg

Lane 5:

HepG2 lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 80 kDa

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• WB

Supplier Data

Western blot - Anti-SUN2 antibody [EPR6557] (AB124916)

All lanes:

Western blot - Anti-SUN2 antibody [EPR6557] (ab124916) at 1/5000 dilution

Lane 1:

HeLa cell Lysate at 20 µg

Lane 2:

Jurkat cell Lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 80 kDa

false

• WB

Supplier Data

Western blot - Anti-SUN2 antibody [EPR6557] (AB124916)

All lanes:

Western blot - Anti-SUN2 antibody [EPR6557] (ab124916) at 1/5000 dilution

All lanes:

Mouse heart lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 80 kDa

false

• WB

Supplier Data

Western blot - Anti-SUN2 antibody [EPR6557] (AB124916)

All lanes:

Western blot - Anti-SUN2 antibody [EPR6557] (ab124916) at 1/5000 dilution

All lanes:

Rat brain lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 80 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-SUN2 antibody [EPR6557] (AB124916)

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.

• WB

CiteAb

Western blot - Anti-SUN2 antibody [EPR6557] (AB124916)

SUN2 western blot using anti-SUN2 antibody [EPR6557] ab124916. Publication image and figure legend from Shaiken, T. E. & Opekun, A. R., 2014, Sci Rep, PubMed 24815916.

ab124916 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab124916 please see the product overview.

Patterns of protein distribution in HeLa cells.Cyt. Cont is a control for cytosolic proteins obtained with the 0.3% Chaps buffer cell lysis (far left panel of bands) for the CDS method; Nuc. Cont is a control for nuclear proteins obtained with the classical method of nuclei isolation in hypotonic buffer (far right panel of bands) for the CDS method. (A) Proteins of cytosol : proteins extracted by regular lysis buffer and Buffer A from the cytoplasm. They are not detected in the perinuclear and the nuclear fractions. Nuclear fractions were obtained with new and classical nuclei extraction techniques (B) Proteins detected in the cytosol and the perinuclear fraction : proteins were detected as cytosolic proteins with both cellular lysis technique; in addition, these proteins also appeared in the perinuclear fraction by extraction with Buffer B. (C) Proteins of perinuclear fraction : proteins are detected only in perinuclear fraction by buffer B extraction. p53 protein was detected with long exposure. (D) Proteins detected in the nuclear and the perinuclear fractions : transcription factor CREB appeared in both fractions. (E) Nuclear proteins : proteins were detected in nuclear fraction. Nuclear proteins were obtained with new and classical nuclei isolation techniques. The PVDF membranes were cropped into two halves and the high and low molecular weight proteins were shown correspondingly.

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