Anti-RBX1 antibody [EPR20185]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBX1 antibody [EPR20185] (AB221548)

Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling RBX1 with ab221548 at 1/4000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear and cytoplasmic staining on human bladder carcinoma tissue sections. The staining pattern observed is consistent with what has been described in the literature (PMID : 23667514).

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)

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Flow Cytometry (Intracellular) - Anti-RBX1 antibody [EPR20185] (AB221548)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling RBX1 with ab221548 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (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) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBX1 antibody [EPR20185] (AB221548)

Immunohistochemical analysis of paraffin-embedded human lung and lung carcinoma tissues labeling RBX1 with ab221548 at 1/4000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Weak nuclear staining of RBX1 on the epithelium cells of human lung (left) compared to strong staining in human lung carcinoma (right).

As documented in the literature ROC1 has higher expression level in cancerous tissue compared to normal tissue (PMID : 19509229).

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-RBX1 antibody [EPR20185] (AB221548)

Immunohistochemical analysis of paraffin-embedded human gastric carcinoma tissue labeling RBX1 with ab221548 at 1/4000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear and cytoplasmic staining on human gastric carcinoma tissue sections. The staining pattern observed is consistent with what has been described in the literature (PMID : 24292229).

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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RBX1 antibody [EPR20185] (AB221548)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling RBX1 with ab221548 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weak cytoplasmic staining on HeLa cell line.

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

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBX1 antibody [EPR20185] (AB221548)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling RBX1 with ab221548 at 1/4000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining is observed on human colon tissue section.

As documented in the literature RBX1 has lower expression level in normal tissue compared to cancerous tissue (PMID : 19509229).

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-RBX1 antibody [EPR20185] (AB221548)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue labeling RBX1 with ab221548 at 1/4000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear and cytoplasmic staining is observed on human colon carcinoma tissue sections.

As documented in the literature RBX1 has higher expression level in cancerous tissue compared to normal tissue (PMID : 19509229).

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.

• IP

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Immunoprecipitation - Anti-RBX1 antibody [EPR20185] (AB221548)

RBX1 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab221548 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab221548 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

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

Lane 2 : ab221548 IP in HeLa whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab221548 in HeLa whole cell lysate.

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

Exposure time : 30 seconds.

The molecular weight observed is consistent with what has been described in the literature (PMID : 22822056).

All lanes:

Immunoprecipitation - Anti-RBX1 antibody [EPR20185] (ab221548)

Predicted band size: 12 kDa

false

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-RBX1 antibody [EPR20185] (AB221548)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed 90% methanol permeabilized NIH/3T3 (mouse embyro fibroblast cell line) cell line labeling RBX1 with ab221548 at 1/60 dilution (red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (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) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBX1 antibody [EPR20185] (AB221548)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling RBX1 with ab221548 at 1/4000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear and cytoplasmic staining on rat colon tissue sections.

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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RBX1 antibody [EPR20185] (AB221548)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embyro fibroblast cell line) cells labeling RBX1 with ab221548 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weak cytoplasmic staining on NIH/3T3 cell line.

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

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBX1 antibody [EPR20185] (AB221548)

Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling RBX1 with ab221548 at 1/4000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear and cytoplasmic staining on mouse stomach tissue sections.

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

Supplier Data

Western blot - Anti-RBX1 antibody [EPR20185] (AB221548)

Exposure time : Lanes 1-3 : 30 seconds; Lane 4 : 10 seconds.

Blocking/Dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 22822056).

All lanes:

Western blot - Anti-RBX1 antibody [EPR20185] (ab221548) at 1/5000 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate at 20 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: 12 kDa

Observed band size: 11 kDa,12 kDa

true

• WB

Supplier Data

Western blot - Anti-RBX1 antibody [EPR20185] (AB221548)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-RBX1 antibody [EPR20185] (ab221548) at 1/1000 dilution

Lane 1:

Human fetal heart lysate at 20 µg

Lane 2:

Human fetal kidney lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/4000 dilution

Predicted band size: 12 kDa

Observed band size: 12 kDa

true

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-RBX1 antibody [EPR20185] (AB221548)

Exposure time : Lanes 1-6 : 3 seconds; Lanes 7-8 : 5 seconds.

Blocking/Dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 22822056).

All lanes:

Western blot - Anti-RBX1 antibody [EPR20185] (ab221548) at 1/1000 dilution

Lane 1:

Mouse heart lysate at 10 µg

Lane 2:

Mouse kidney lysate at 10 µg

Lane 3:

Mouse spleen lysate at 10 µg

Lane 4:

Rat brain lysate at 10 µg

Lane 5:

Rat kidney lysate at 10 µg

Lane 6:

Rat spleen lysate at 10 µg

Lane 7:

RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 8:

PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

Lane 9:

NIH/3T3 (mouse embyro fibroblast cell line) 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: 12 kDa

Observed band size: 11 kDa,12 kDa

true