Anti-Fukutin antibody [EPR7913]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fukutin antibody [EPR7913] (AB131280)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling Fukutin with ab131280 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Unknown

Western blot - Anti-Fukutin antibody [EPR7913] (AB131280)

All lanes:

Western blot - Anti-Fukutin antibody [EPR7913] (ab131280) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

A549 cell lysate at 10 µg

Lane 3:

Fetal heart tissue lysate at 10 µg

Lane 4:

BxPC-3 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 53 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Fukutin antibody [EPR7913] (AB131280)

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-Fukutin antibody [EPR7913] (AB131280)

Fukutin western blot using anti-Fukutin antibody [EPR7913] ab131280. Publication image and figure legend from Gerin, I., Ury, B., et al., 2016, Nat Commun, PubMed 27194101.

ab131280 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 ab131280 please see the product overview.

FKTN and FKRP can transfer ribitol phosphate to recombinant α-dystroglycan.(a) Schematic representation of α-dystroglycan fragments used in this study. (b) Transfer of [3H]-ribitol from CDP-[1-3H]-ribitol to α-dystroglycan (~1 μg) by recombinant FKTN and FKRP (~0.1 and ~0.4 μg, respectively). After incubation for 16 h at 30 °C, proteins were recovered using streptavidin beads and incorporated radioactivity was determined with a scintillation counter. Values are means±s.d. of three biological replicates from one representative experiments out of three. Asterisks indicate p<0.05 in Student's t-test. (c) Incorporation of [32P] from [32P]-CDP-ribitol to different fragments of α-dystroglycan by FKTN and FKRP. After 16 h of incubation at 30 °C, proteins were incubated with (+) or without (-) alkaline phosphatase (AP) and resolved by SDS–PAGE. 32P was detected with a storage phosphorimager. (d) Effect of deglycosylation of α-dystroglycan (aa 1–485) with a 'deglycosylation mix' on the incorporation of radioactivity from [32P]-CDP-ribitol catalysed by FKRP or FKTN. The deglycosylation treatment of α-dystroglycan performed either before (two left panels) or after (two right panels) ribitol-phosphorylation decreased the apparent molecular weight of α-dystroglycan, but did not reduce the incorporated radioactivity.

false