Anti-ACTR1B antibody [EPR16969]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ACTR1B antibody [EPR16969] (AB203835)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling ACTR1B with ab203835 at 1/250 dilution followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab203835 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACTR1B antibody [EPR16969] (AB203835)

Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling ACTR1B with ab203835 at 1/300 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on Human cervix carcinoma tissue 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) (ab97051) at 1/500 dilution.

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

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ACTR1B antibody [EPR16969] (AB203835)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling ACTR1Bwith ab203835 at 1/300 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ACTR1B antibody [EPR16969] (AB203835)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling ACTR1B with ab203835 at 1/250 dilution followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on MCF7 cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab203835 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.

-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• IP

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Immunoprecipitation - Anti-ACTR1B antibody [EPR16969] (AB203835)

ACTR1B was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab203835 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab203835 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution.

Lane 1 : HeLa whole cell lysate 10ug (Input). Lane 2 : ab203835 IP in HeLa whole cell lysate. Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab203835 in HeLa whole cell lysate.

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

Exposure time : 5 seconds.

All lanes:

Immunoprecipitation - Anti-ACTR1B antibody [EPR16969] (ab203835)

Predicted band size: 42 kDa

false

• WB

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Western blot - Anti-ACTR1B antibody [EPR16969] (AB203835)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ACTR1B antibody [EPR16969] (ab203835) at 1/10000 dilution

Lane 1:

MOLT-4 (Human lymphoblastic leukemia cell line) whole cell lysate at 10 µg

Lane 2:

HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 10 µg

Lane 3:

293 (Human epithelial cells from embryonic kidney) whole cell lysate at 10 µg

Lane 4:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-ACTR1B antibody [EPR16969] (AB203835)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ACTR1B antibody [EPR16969] (ab203835) at 1/10000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human fetal kidney lysate at 10 µg

Lane 3:

Human fetal spleen lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-ACTR1B antibody [EPR16969] (AB203835)

Blocking/Dilution buffer : 5% NFDM/TBST.

Lane 1:

Western blot - Anti-ACTR1B + ACTR1A antibody [EPR16968(B)] (<a href='/en-us/products/primary-antibodies/actr1b-actr1a-antibody-epr16968b-ab203833'>ab203833</a>) at 1/1000 dilution

Lane 1:

Western blot - Anti-ACTR1B antibody [EPR16969] (ab203835) at 1/1000 dilution

All lanes:

Full length Human ACTR1A recombination protein at 0.01 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: 42 kDa

Observed band size: 42 kDa

false

Exposure time: 3s