Anti-SFPQ 抗体

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-SFPQ antibody (AB38148)

Immunofluorescence analysis of mouse amygdala tissue, staining SFPQ (green) with ab38148 at 1/200 dilution. An AlexaFluor®488-conjugated anti-rabbit IgG was used as the secondary antibody.

Image from Ke Y et al., PLoS One. 2012;7(4):e35678. Epub 2012 Apr 25. Fig 2.; doi:10.1371/journal.pone.0035678; April 25, 2012, PLoS ONE 7(4): e35678.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SFPQ antibody (AB38148)

IHC image of SFPQ staining in Human colon cancer formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab38148, 1μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SFPQ antibody (AB38148)

ab38148 staining SFPQ in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-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 ab38148 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 100% methanol (5 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• WB

Project

Western blot - Anti-SFPQ antibody (AB38148)

We are unsure as to the identity of the 90kDa band as no isoforms at this molecular weight are known. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

All lanes:

Western blot - Anti-SFPQ antibody (ab38148) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

Lane 4:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 76 kDa

Observed band size: 90 kDa,95 kDa

false

• WB

Unknown

Western blot - Anti-SFPQ antibody (AB38148)

All lanes:

Western blot - Anti-SFPQ antibody (ab38148) at 1 µg/mL

All lanes:

NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 76 kDa

Observed band size: 100 kDa

true

Exposure time: 1min

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-SFPQ antibody (AB38148)

Immunocytochemistry-immunofluorescence using Anti-SFPQ antibody, ab38148. Publication image from Li, X. J. et al., 2015, Mol Neurodegener, 26334913. Legend direct from paper.

Cytoplasmic mislocalization of PSF, reduction of NMHC II-B, and abnormal splicing of NMHC II-B in the brains of ALS patients. a Double immunofluorescent staining of the cortex and spinal cord from the ALS patient and non-ALS individual (control) with antibodies to PSF and NeuN. Scale bars : cortex : 10 µm. Spinal cord : 50 µm. Note that ALS patient brain shows cytoplasmic distribution of PSF and NeuN. b Immunostaining of the brain cortex in control and ALS patients with single anti-PSF only revealed PSF immunofluorescent staining. c Quantitative analysis of the percentage of cells with cytoplasmic PSF and NeuN staining. The staining of the nuclear protein TFIIB served as controls. The data are mean ± SE by counting 60 to 80 cells per sample (n = 3per group). *p < 0.05, **p < 0.01, ***p < 0.001

• WB

CiteAb

Western blot - Anti-SFPQ antibody (AB38148)

Western Blotting using Anti-SFPQ antibody, ab38148. Publication image from Li, X. J. et al., 2015, Mol Neurodegener, 26334913. Legend direct from paper.

Interactions of TDP-43 with PSF and reduced NMCH II-B in ALS patient brains. a Representative images of cytoplasmic mislocalization of PSF and TDP-43 in the ALS patient brain. Double immunofluorescent staining of the brain cortex from ALS patient and non-ALS individual with antibodies to TDP-43 and PSF. Note that ALS patient brain contains cytoplasmic TDP-43 and PSF. Scale bar : 10 µm. b Immunoprecipitation of TDP-43 in the cortex tissue of ALS patient showing the presence of TDP-43, PSF, and NeuN in the cytoplasmic fraction. CTL is the sample from a normal individual. Rabbit IgG served as a control immunoprecipitation. Whole : cortex total lysates; Cyto : cytoplasm of cortex lysates; Nucl : nuclear of cortex lysates; c The ratios (mean ± SE, n = 3) of immunoprecipitate (IP) to input and cytoplasmic or nuclear proteins to total lysates on western blots are presented. *p < 0.05; **p < 0.01 compared with CTL. d RT-PCR gel result (the upper panel) and quantification of the relative levels (the lower panel) of +N30 and -N30 isoforms in the brain cortex samples from 3 normal individuals (control), 3 Alzheimer’s disease (AD), and 5 ALS patients. e NMHC II-B western blot analysis of the brain cortex from normal individuals, Alzheimer’s disease (AD) and 5 ALS patients showing decreased expression of NMCH II-B in ALS patient brains

false

• WB

CiteAb

Western blot - Anti-SFPQ antibody (AB38148)

Western Blotting using Anti-SFPQ antibody, ab38148. Publication image from Li, X. J. et al., 2015, Mol Neurodegener, 26334913. Legend direct from paper.

Interactions of TDP-43 with PSF and NeuN. a Representative western blots showing co-immunoprecipitation of transfected flag-TDP-43 (WT or mutant TDP-43 (M337V)) by anti-flag and endogenous PSF, NeuN in mouse N2A cells. IgG immunoprecipitation served as controls. The ratios of precipitated to input were obtained from 3 independent experiments and shown beneath the blots. b The in vivo interaction of TDP-43 (M337V) with PSF and NeuN in the non-transgenic (C) pig and transgenic TDP-43 (TG) pig brain. The immunoprecipitation was also pre-treated with RNase to remove RNA. An antibody to TDP-43 (12892-1-AP from Proteintech) selectively detected human TDP-43, but not endogenous pig TDP-43. The interaction of FUS with TDP-43, which depends on RNA, served as a control. Quantitative data (ratio of IP/input) are presented beneath the blots. IP bands indicated by arrows were used for quantification. *p < 0.05, **p < 0.01 and ***p < 0.001 compared with –RNase treatment

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