Anti-RRM1 antibody [EPR8483]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RRM1 antibody [EPR8483] (AB137114)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling RRM1 with purified ab137114 at 1/1000 dilution (0.95 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• ICC/IF

AbReview32463****

Immunocytochemistry/ Immunofluorescence - Anti-RRM1 antibody [EPR8483] (AB137114)

Unpurified ab137114 (1/200) staining RRM1 in Hela cells (green). Cells were fixed in paraformaldehyde, permeabilised in 0.5%Triton X-100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.

Image courtesy of an Abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-RRM1 antibody [EPR8483] (AB137114)

unpurified ab137114 staining RRM1 in the human cell line HeLa (human cervix adenocarcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde, permiabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control : Rabbit monoclonal IgG (Black)

Unlabelled control : Cell without incubation with primary antibody and secondary antibody (Blue)

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RRM1 antibody [EPR8483] (AB137114)

Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labelling RRM1 with unpurified ab137114 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RRM1 antibody [EPR8483] (AB137114)

Immunocytochemistry/ Immunofluorescence analysis of HT-29 (Human colorectal adenocarcinoma epithelial cell) cells labeling RRM1 with purified ab137114 at 1/100 dilution (9.5 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RRM1 antibody [EPR8483] (AB137114)

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labelling RRM1 with ab137114 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-RRM1 antibody [EPR8483] (AB137114)

All lanes:

Western blot - Anti-RRM1 antibody [EPR8483] (ab137114) at 1/10000 dilution

Lane 1:

HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 2:

A549 (Human lung carcinoma epithelial cell) whole cell lysates at 20 µg

Lane 3:

Mouse colon lysates at 20 µg

Lane 4:

Rat brain lysates at 20 µg

Lane 5:

Rat colon lysates 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 1/20000 dilution

Predicted band size: 90 kDa

Observed band size: 90 kDa

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• WB

Unknown

Western blot - Anti-RRM1 antibody [EPR8483] (AB137114)

All lanes:

Western blot - Anti-RRM1 antibody [EPR8483] (ab137114) at 1/10000 dilution

Lane 1:

HT29 cell lysate at 10 µg

Lane 2:

A549 cell lysate at 10 µg

Lane 3:

MCF7 cell lysate at 10 µg

Lane 4:

HeLa cell lysate at 10 µg

Secondary

All lanes:

Standard HRP-labeled goat anti-rabbit at 1/2000 dilution

Predicted band size: 90 kDa

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• WB

CiteAb

Western blot - Anti-RRM1 antibody [EPR8483] (AB137114)

RRM1 western blot using anti-RRM1 antibody [EPR8483] ab137114. Publication image and figure legend from Zhou, J., Zhang, L., et al., 2020, Cancer Med, PubMed 31823522.

ab137114 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab137114 please see the product overview.

Expression of RRM1, STIM1, and TRIM21 was associated with the level of acquired gemcitabine (GEM) resistance. A, Venn diagram showed the top 20 consistently upregulated mRNAs in both GEM-resistant cell lines. B, Six intersected genes selected from among the top 20 consistently upregulated mRNAs. C and D, Total RNA and cytoplasmic proteins were extracted from BxPC-3, BxPC-3-GR, CFPAC-1, and CFPAC-1-GR. mRNA and protein expression levels of RRM1, STIM1, and TRIM21 were determined by quantitative reverse-transcription polymerase chain reaction and western blot analysis. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a control. E and F, RRM1, STIM1, and TRIM21 protein expression levels were measured by western blot analysis in both parental cell lines and their derived GEM subclones. GAPDH was used as a control. G, RRM1, STIM1, and TRIM21 protein expression levels among normal pancreatic and pancreatic cancer (PC) cell lines were evaluated by western blot analysis. GAPDH was used as a control. H, Histogram shows IC50 values for GEM in AsPC-1, BxPC-3, CFPAC-1, MIA PaCa-2, and PANC-1 cell lines. **p < .01, ***p < .001; comparisons indicated by lines

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