Anti-ARL8A antibody [EPR24376-2]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARL8A antibody [EPR24376-2] (AB270979)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labelling ARL8A with ab270979 at 1/50 (10.02 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab270979 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Cytoplasmic staining on human placenta (PMID : 15331635).

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

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-ARL8A antibody [EPR24376-2] (AB270979)

ARL8A was immunoprecipitated from 0.35 mg SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate 10 μg with ab270979 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270979 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : SH-SY5Y whole cell lysate 10 μg

Lane 2 : ab270979 IP in SH-SY5Y whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab270979 in SH-SY5Y whole cell lysate

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

Exposure time : 76 seconds

This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Immunoprecipitation - Anti-ARL8A antibody [EPR24376-2] (ab270979)

Predicted band size: 21 kDa

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• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARL8A antibody [EPR24376-2] (AB270979)

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labelling ARL8A with ab270979 at 1/50 (10.02 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab270979 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Cytoplasmic staining on rat cerebrum.

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

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ARL8A antibody [EPR24376-2] (AB270979)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a cells labelling ARL8A with ab270979 at 1/50 (10.02 μg/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in Neuro-2a cell line.

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 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 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARL8A antibody [EPR24376-2] (AB270979)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labelling ARL8A with ab270979 at 1/50 (10.02 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab270979 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Cytoplasmic staining on mouse cerebrum.

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

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-ARL8A antibody [EPR24376-2] (AB270979)

ARL8A was immunoprecipitated from 0.35 mg Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate 10 μg with ab270979 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270979 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Neuro-2a whole cell lysate 10 μg

Lane 2 : ab270979 IP in Neuro-2a whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab270979 in Neuro-2a whole cell lysate

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

Exposure time : 58 seconds

This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Immunoprecipitation - Anti-ARL8A antibody [EPR24376-2] (ab270979)

Predicted band size: 21 kDa

false

• WB

Lab

Western blot - Anti-ARL8A antibody [EPR24376-2] (AB270979)

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

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-ARL8A antibody [EPR24376-2] (ab270979) at 1/1000 dilution

Lane 1:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 40 µg

Lane 2:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 40 µg

Lane 3:

SK-BR-3 (human breast adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 4:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 40 µg

Lane 6:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 40 µg

Lane 7:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 40 µg

Lane 8:

C6 (rat glial tumor glial cell) 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 1/50000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false

• WB

Lab

Western blot - Anti-ARL8A antibody [EPR24376-2] (AB270979)

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

Lanes 1-2 : This blot was developed using a higher sensitivity ECL substrate.

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-ARL8A antibody [EPR24376-2] (ab270979) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 40 µg

Lane 2:

Human placenta tissue lysate at 40 µg

Lane 3:

Mouse brain tissue lysate at 40 µg

Lane 4:

Mouse placenta tissue lysate at 40 µg

Lane 5:

Rat brain tissue lysate at 40 µg

Lane 6:

Rat placenta tissue lysate at 40 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 1/1000 dilution

Lanes 3 - 6:

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 1/50000 dilution

Predicted band size: 21 kDa

false

• WB

Lab

Western blot - Anti-ARL8A antibody [EPR24376-2] (AB270979)

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

This antibody reacts with ARL8A, but not ARL8B.

Exposure time : 3 seconds.

All lanes:

Western blot - Anti-ARL8A antibody [EPR24376-2] (ab270979) at 1/1000 dilution

Lane 1:

His-tagged human ARL8A recombinant protein at 0.02 µg

Lane 2:

His-tagged human ARL8B recombinant protein at 0.02 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: 21 kDa

false