Anti-Dynamin 2 antibody [EPR9053(2)(ABC)]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dynamin 2 antibody [EPR9053(2)(ABC)] (AB151555)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling Dynamin 2 with ab151555 at 1/50 dilution.

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dynamin 2 antibody [EPR9053(2)(ABC)] (AB151555)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labelling Dynamin 2 with ab151555 at 1/50 dilution.

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

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Dynamin 2 antibody [EPR9053(2)(ABC)] (AB151555)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized K-562 cells labelling Dynamin 2 with ab151555 at 1/50 (19.56 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in K-562 cell line ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 2 ug/ml dilution.

• WB

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Western blot - Anti-Dynamin 2 antibody [EPR9053(2)(ABC)] (AB151555)

All lanes:

Western blot - Anti-Dynamin 2 antibody [EPR9053(2)(ABC)] (ab151555) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

K562 cell lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Lane 4:

Fetal brain tissue lysate at 10 µg

Predicted band size: 98 kDa

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• WB

Lab

Western blot - Anti-Dynamin 2 antibody [EPR9053(2)(ABC)] (AB151555)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 36369230).

All lanes:

Western blot - Anti-Dynamin 2 antibody [EPR9053(2)(ABC)] (ab151555) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 3:

Mouse testis tissue lysate at 20 µg

Lane 4:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 6:

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

Lane 7:

Rat thymus tissue lysate at 20 µg

Lane 8:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 9:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate 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

Observed band size: 98-100 kDa

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Exposure time: 180s

• WB

CiteAb

Western blot - Anti-Dynamin 2 antibody [EPR9053(2)(ABC)] (AB151555)

Dynamin 2 western blot using anti-Dynamin 2 antibody [EPR9053(2)(ABC)] ab151555. Publication image and figure legend from Li, M., Zhang, D., et al., 2020, Front Microbiol, PubMed 32210929.

ab151555 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 ab151555 please see the product overview.

Zika virus (ZIKV) entry depends on dynamin II. (A) Cell viability upon dynasore treatment was assessed by the MTT assay. (B,C) Treatment with dynasore inhibited the ZIKV entry into T98G cells. T98G cells treated with dynasore were incubated with ZIKV at a multiplicity of infection (MOI) of 10 for 1 h at 4°C and then shifted to 37°C for 1 h. The internalized viruses were determined by RT-qPCR (B) and infectious center assay (C). (D) To verify dynamin II knockdown, protein samples from the cells transfected with each siRNA were analyzed by immunoblot for dynamin II. (E,F) T98G cells were transfected with siRNA targeting dynamin II or control siRNA for 48 h, followed by infection with ZIKV at an MOI of 10 for 1 h at 4°C and then shifted to 37°C for 1 h. The internalized viruses were determined by RT-qPCR (E) and infectious center assay (F). (G) T98G cells were transfected with siRNA targeting dynamin II or control siRNA for 48 h and then incubated with 10 μg/ml AF-555-labeled transferrin for 10 min at 37°C. The cells were fixed and stained with DAPI. Scale bars in all panels represent 10 μm. (H) T98G cells were transfected with siRNA targeting dynamin II or control siRNA for 48 h, followed by infection with ZIKV at an MOI of 0.5. At 48 h post-infection, the cells were fixed and analyzed under an Olympus microscope. Scale bars in all panels represent 50 μm. (I) The cells transfected with the dynamin II wild-type or dominant-negative plasmid construct were infected with ZIKV at an MOI of 0.5. At 48 h post-infection, the cells were analyzed by flow cytometry. One representative experiment out of three is shown (D). Representative confocal images from three independent experiments are shown (G,H). The data shown are the mean ± SD of three independent experiments (A–C,E,F,I). *p < 0.05; **p < 0.01; ***p < 0.001.

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