Anti-Visfatin antibody [EPR21984]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Visfatin antibody [EPR21984] (AB236873)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HCT 116 (human colorectal carcinoma cell line) cell line labeling Visfatin with ab236873 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Visfatin antibody [EPR21984] (AB236873)

Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling Visfatin with ab236873 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear and cytoplasmic staining in human bladder cancer (PMID : 26396920) is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Visfatin antibody [EPR21984] (AB236873)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling Visfatin with ab236873 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear and cytoplasmic staining in human breast cancer(PMID : 21784959) is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Visfatin antibody [EPR21984] (AB236873)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized K562 (human chronic myelogenous leukemia cell line from bone marrow) cell line labeling Visfatin with ab236873 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody

• IP

Supplier Data

Immunoprecipitation - Anti-Visfatin antibody [EPR21984] (AB236873)

Visfatin was immunoprecipitated from 0.35 mg HCT 116 (human colorectal carcinoma cell line) whole cell lysate with ab236873 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab236873 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : HTC 116 whole cell lysate 10 μg (Input).
Lane 2 : ab236873 IP in HCT 116 whole cell lysate (+).
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab236873 in HCT 116 whole cell lysate (-).

Blocking/Dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-Visfatin antibody [EPR21984] (ab236873)

Predicted band size: 55 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Visfatin antibody [EPR21984] (AB236873)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Visfatin with ab236873 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear and cytoplasmic staining in mouse kidney (PMID : 29104634; PMID : 24287278) is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Visfatin antibody [EPR21984] (AB236873)

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Visfatin with ab236873 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear and cytoplasmic staining in rat liver (PMID : 29104634; PMID : 24603648) is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

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

• WB

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Western blot - Anti-Visfatin antibody [EPR21984] (AB236873)

Exposure time : Lanes 1-3 : 37 seconds; Lanes 4-5 : 8 seconds; Lane 6 : 26 seconds.

Blocking and diluting buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 25368373, PMID : 12782293).

All lanes:

Western blot - Anti-Visfatin antibody [EPR21984] (ab236873) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 2:

HT1080 (human fibrosarcoma cell line) whole cell lysate at 20 µg

Lane 3:

PC-3 (human prostate adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

NCI-H460 (human lung cancer cell line) whole cell lysate at 20 µg

Lane 5:

Human breast cancer lysate at 20 µg

Lane 6:

K562 (human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 10 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 1/100000 dilution

Predicted band size: 55 kDa

Observed band size: 56 kDa

false

• WB

Supplier Data

Western blot - Anti-Visfatin antibody [EPR21984] (AB236873)

Blocking and diluting buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 25368373, PMID : 12782293).

All lanes:

Western blot - Anti-Visfatin antibody [EPR21984] (ab236873) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

HCT 116 (human colorectal carcinoma cell line) whole cell lysate at 20 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 20 µg

Lane 4:

C2C12 (mouse myoblast cell line) whole cell lysate at 20 µg

Lane 5:

Human fetal brain lysate at 10 µg

Lane 6:

Human heart lysate at 10 µg

Lane 7:

Human fetal kidney lysate at 10 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 1/100000 dilution

Predicted band size: 55 kDa

Observed band size: 56 kDa

false

Exposure time: 3min