Anti-IL-16 antibody [EPR19988]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IL-16 antibody [EPR19988] (AB207181)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed H9 (Human cutaneous T lymphocyte lymphoma cell line) cells labeling IL-16with ab207181 at 1/600 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluorr^® 488) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-16 antibody [EPR19988] (AB207181)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling IL-16 with ab207181 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on stromal cells of human colon is observed [PMID : 11709514]. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IL-16 antibody [EPR19988] (AB207181)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized H9 (Human cutaneous T lymphocyte lymphoma cell line) cells labeling IL-16 with ab207181 at 1/100 dilution followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on H9 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-16 antibody [EPR19988] (AB207181)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling IL-16 with ab207181 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human tonsil is observed [PMID : 10946273]. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

• WB

Supplier Data

Western blot - Anti-IL-16 antibody [EPR19988] (AB207181)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1 : 5 seconds, Lane 2 : 15 seconds, Lane 3 : 3 minutes.

The band at 80 kDa is pro-IL-16 and the bands at 40-75 kDa are cleaved fragments. This is consistent with what has been described in the literature (PMID : 15187155, PMID : 9144227, PMID : 9743378, PMID : 14734747).

All lanes:

Western blot - Anti-IL-16 antibody [EPR19988] (ab207181) at 1/1000 dilution

Lane 1:

Human thymus tissue lysate at 10 µg

Lane 2:

Human spleen tissue lysate at 10 µg

Lane 3:

Human tonsil tissue lysate at 20 µg

Secondary

Lanes 1 - 2:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/4000 dilution

Lane 3:

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: 142 kDa

Observed band size: 75-40 kDa,80 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-IL-16 antibody [EPR19988] (AB207181)

IL-16 was immunoprecipitated from 0.35 mg of H9 (Human cutaneous T lymphocyte lymphoma cell line) whole cell lysate with ab207181 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab207181 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : H9 whole cell lysate, 10 μg (Input).

Lane 2 : ab207181 IP in H9 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab207181 in H9 whole cell lysate.

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

Exposure time : 10 seconds.

Note : The band at around 50kDa is a cleaved form of IL-16.

All lanes:

Immunoprecipitation - Anti-IL-16 antibody [EPR19988] (ab207181)

Predicted band size: 142 kDa

Observed band size: 37-80 kDa

false

• WB

Supplier Data

Western blot - Anti-IL-16 antibody [EPR19988] (AB207181)

Blocking/Dilution buffer : 5% NFDM/TBST.

The band at 80 kDa is pro-IL-16 and the bands at 40-75 kDa are cleaved fragments. This is consistent with what has been described in the literature (PMID : 15187155, PMID : 9144227, PMID : 9743378, PMID : 14734747).

All lanes:

Western blot - Anti-IL-16 antibody [EPR19988] (ab207181) at 1/1000 dilution

Lane 1:

H9 (Human cutaneous T lymphocyte lymphoma cell line) whole cell lysate at 10 µg

Lane 2:

Jurkat (Human T cell leukemia cell line from peripheral blood) 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: 142 kDa

Observed band size: 75-40 kDa,80 kDa

false

Exposure time: 3min