Anti-IL-17A antibody [RM1202] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-17A antibody [RM1202] - BSA and Azide free (AB318151)

This data was developed using ab318150, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse skin tissue labeling IL-17A with ab318150 at 1/1000 (0.498 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse skin. The section was incubated with ab318150 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-17A antibody [RM1202] - BSA and Azide free (AB318151)

This data was developed using ab318150, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) EL4 (mouse lymphoma T lymphocyte) treated with 10 nM PMA, 1uM Ionomycin and 300ng/ml BFA for 24 hours cell pellet. (B) Untreated EL4 cell pellet. tissue labeling IL-17A with ab318150 at 1/1000 (0.498 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) EL4 treated with 10 nM PMA, 1uM Ionomycin and 300ng/ml BFA for 24 hours cell pellet, weak staining on (B) untreated EL4 cell pellet. The section was incubated with ab318150 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IL-17A antibody [RM1202] - BSA and Azide free (AB318151)

This data was developed using ab318150, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized EL4 (mouse lymphoma T lymphocyte) cells labelling IL-17A with ab318150 at 1/50 (9.96 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing weak staining in EL4 cell line (shown in green). The expression increased after treatment with PMA (50 ng/ml), lonomycin Ca2 + Salt (500 ng/ml) and Brefeldin A (500 ng/ml) overnight (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-IL-17A antibody [RM1202] - BSA and Azide free (AB318151)

IL-17A was immunoprecipitated from 0.35 mg EL4 (mouse lymphoma T lymphocyte) treated with 10nM PMA, 1uM Ionomycin and 300ng/ml BFA for 24h, whole cell lysate with ab318150 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab318150 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : EL4 (mouse lymphoma T lymphocyte) treated with 10nM PMA, 1uM Ionomycin and 300ng/ml BFA for 24h, whole cell lysate
Lane 2 : ab318150 IP in EL4 (mouse lymphoma T lymphocyte) treated with 10nM PMA, 1uM Ionomycin and 300ng/ml BFA for 24h, whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab318150 in treated with 10nM PMA, 1uM Ionomycin and 300ng/ml BFA for 24h, whole cell lysate

All lanes:

Immunoprecipitation - Anti-IL-17A antibody [RM1202] (<a href='/en-us/products/primary-antibodies/il-17a-antibody-rm1202-ab318150'>ab318150</a>) at 1/30 dilution

All lanes:

EL4 (mouse lymphoma T lymphocyte) treated with 10nM PMA, 1uM Ionomycin and 300ng/ml BFA for 24h, whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 145s

• WB

Supplier Data

Western blot - Anti-IL-17A antibody [RM1202] - BSA and Azide free (AB318151)

This data was developed using ab318150, the same antibody clone in a different buffer formulation.

The expression profile is consistent with the literature (PMID : 17355969).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-IL-17A antibody [RM1202] (<a href='/en-us/products/primary-antibodies/il-17a-antibody-rm1202-ab318150'>ab318150</a>) at 1/1000 dilution

Lane 1:

Untreated NR8383 (rat alveolar macrophage) whole cell lysate at 40 µg

Lane 2:

NR8383 treated with 50ng/ml PMA, 500ng/ml lonomycin Ca2+Salt and 500ng/ml BFA overnight, whole cell lysate at 40 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: 25 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-IL-17A antibody [RM1202] - BSA and Azide free (AB318151)

This data was developed using ab318150, the same antibody clone in a different buffer formulation.

Expression of IL-17A can be induced by PMA and Ionomycin treatment (PMID 28382171).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-IL-17A antibody [RM1202] (<a href='/en-us/products/primary-antibodies/il-17a-antibody-rm1202-ab318150'>ab318150</a>) at 1/1000 dilution

Lane 1:

Untreated EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 40 µg

Lane 2:

EL4 treated with 10 nM PMA, 1uM Ionomycin and 300ng/ml BFA for 24 hours, whole cell lysate at 40 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: 14-25 kDa,36 kDa

false

Exposure time: 81s

• I-ELISA

Supplier Data

Indirect ELISA - Anti-IL-17A antibody [RM1202] - BSA and Azide free (AB318151)

This data was developed using ab318150, the same antibody clone in a different buffer formulation.

Indirect ELISA analysis of ab318150 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution.

Antigen : Mouse IL-17A,Mouse IL-17F.

Antigen concentration : 1000 ng/ml

This antibody does not cross-react with mouse IL-17F.