rabbit anti agr2 monoclonal antibody  (Cell Signaling Technology Inc)

Journal: Molecular and cellular biochemistry

Article Title: Agr2 in cancer and beyond: unraveling its role during protein synthesis, ER stress, and as a predictive biomarker.

doi: 10.1007/s11010-025-05318-8

Figure Lengend Snippet: Fig. 4 Effects of Agr2 on cel- lular function, protein targets, and interaction partners. Agr2 is involved in tumor initiation and dissemination, cellular signaling to mediate different cell-fate decisions, regulating proteosta- sis and protein trafficking, and chemoresistance

Article Snippet: The Agr2 primary antibody, Cat. no. 13062 was obtained from Cell Signaling Technologies.

Techniques:

Journal: Molecular and cellular biochemistry

Article Title: Agr2 in cancer and beyond: unraveling its role during protein synthesis, ER stress, and as a predictive biomarker.

doi: 10.1007/s11010-025-05318-8

Figure Lengend Snippet: Fig. 5 Structure of chemicals or molecules that bind to and inhibit the activity of Agr2. A MG132 is a tri-peptide aldehyde with a mem- brane-permeable proteasome inhibitory function. B Bortezomib is a first-in-class ubiquitin–proteasome inhibitor used to treat multiple myeloma and mantle cell lymphoma. C C-3353 is an aromatic N-sul- phonamide derivative of aziridine-2-carboxylic acid with inhibitory

Article Snippet: The Agr2 primary antibody, Cat. no. 13062 was obtained from Cell Signaling Technologies.

Techniques: Activity Assay, Ubiquitin Proteomics

Journal: Cancer Research Communications

Article Title: High-Throughput Drug Screening of Clear Cell Ovarian Cancer Organoids Reveals Vulnerability to Proteasome Inhibitors and Dinaciclib and Identifies AGR2 as a Therapeutic Target

doi: 10.1158/2767-9764.CRC-25-0024

Figure Lengend Snippet: AGR2-high CCC organoids are sensitive to bortezomib, and AGR2 knockout reduces tumor growth. A, Heatmap showing the IC 50 of drugs selected through the HTDS of 361 compounds of nine organoids (18-015, 19-079, 19-055, 19-076, 18-148, 19-042, 19-001, 19-010, and 19-044). High, green; low, magenta. B, Genes shared in at least two of the UPR-related pathways (GOBP_IRE1_mediated_unfolded_protein_response, GOBP_regulation_of_PERK_mediated_unfolded_protein_response, and GOBP_ATF6_mediated_unfolded_protein_response). AGR2 was significantly downregulated in the bortezomib-resistant 18-015 CCC organoid ( P = 0.0049). C, Read count of AGR2 in RNA sequencing of bortezomib-resistant (18-015) and other CCC organoids. D, Immunoblot analysis of UPR-related genes, including AGR2 in bortezomib-resistant (18-015) and other CCC organoids (19-001, 19-042, 19-055, 19-076, and 19-079). E, Immunoblot analysis of AGR2 -KO CCC organoid (19-042) confirmed knockout of AGR2. F, AGR2 -KO organoids exhibited significantly slower growth than the control ( P = 0.014). G, Cell cycle analysis of the AGR2 -KO CCC organoid (19-042) showed a significant increase in the G 2 –M fraction compared with the control ( P = 0.028).

Article Snippet: Unstained 4-μm sections of clinical specimens were deparaffinized, rehydrated, and incubated with a primary antibody against AGR2 (1:500 dilution, D9V2F, XP Rabbit mAb #13062, Cell Signaling Technology) at room temperature for 30 minutes.

Techniques: Knock-Out, RNA Sequencing, Western Blot, Control, Cell Cycle Assay

Journal: Cancer Research Communications

Article Title: High-Throughput Drug Screening of Clear Cell Ovarian Cancer Organoids Reveals Vulnerability to Proteasome Inhibitors and Dinaciclib and Identifies AGR2 as a Therapeutic Target

doi: 10.1158/2767-9764.CRC-25-0024

Figure Lengend Snippet: AGR2 knockout sensitizes CCC organoids to carboplatin through upregulation of SLFN11 and induces UPR and ER stress. A, Dose–response curve of bortezomib, dinaciclib, and carboplatin in AGR2 -KO organoids. AGR2 -KO organoids showed increased sensitivity to bortezomib ( P = 0.007 and 0.012) and carboplatin (both P = 0.010) but not to dinaciclib. B, Correlation between AGR2 and SLFN11 mRNA in the Cancer Cell Line Encyclopedia dataset (Broad, 2019; ref. ). AGR2 expression was significantly related to low SLFN11 expression ( P = 5.087 × 10 −14 ). C, Immunoblot analysis of UPR pathway genes and SLFN11 in AGR2 -KO CCC organoids. D, Activation of the UPR pathway in AGR2 -KO CCC organoids (Hallmark_unfolded_protein_response, P = 0.0022; GOBP_ATF6_mediated_ unfolded_protein_response, P = 0.00012; GOBP_IRE1_mediated_unfolded_protein_response, P = 0.00073; and GOBP_regulation_of_PERK_mediated_unfolded_protein_response, P = 0.76, compared with the control). E, TEM images the of AGR2 -KO CCC organoid. The arrowhead indicates the ER. AGR2 knockout showed significant ER lumen dilatation (clone B, P < 0.001 and clone C, P < 0.001). F, Higher AGR2 expression in CCC ( n = 51) than in HGSC ( n = 43; P < 0.001). AGR2 expression was not correlated with progression-free survival in CCC ( P = 0.74). G, Schematic representation of the identified mechanisms in this study. CCC exhibits high UPR, which is further upregulated by PIs to induce apoptosis. There are many AGR2-high CCCs, and AGR2 inhibition upregulates UPR and SLFN11 expression, leading to growth inhibition and platinum sensitivity.

Article Snippet: Unstained 4-μm sections of clinical specimens were deparaffinized, rehydrated, and incubated with a primary antibody against AGR2 (1:500 dilution, D9V2F, XP Rabbit mAb #13062, Cell Signaling Technology) at room temperature for 30 minutes.

Techniques: Knock-Out, Expressing, Western Blot, Activation Assay, Control, Inhibition

Journal: Communications biology

Article Title: Human cytomegalovirus UL82 promotes cell cycle progression of colorectal cancer by upregulating AGR2.

doi: 10.1038/s42003-025-07674-z

Figure Lengend Snippet: Fig. 7 | Schematic diagram of the UL82/DDX5 complex upregulating AGR2 and promoting nucleotide synthesis in CRC cells. After HCMV infection, UL82 is released into the nucleus of CRC cells. UL82 then interacts with DDX5 to block its transcriptional repression of AGR2, thereby pro- moting AGR2 expression. The UL82-AGR2 axis enhances nucleotide metabolism by upregulating nucleotide synthesis enzyme levels in colorectal cancer cells.

Article Snippet: Communications Biology | (2025) 8:251 10 The membrane was incubated overnight at 4 °C with primary antibodies against AGR2 (Abclonal, A7064, 1:1000), Flag (Origene, TA180144, 1:2000), DDX5 (Abclonal, A11339, 1:1000), c-Myc (Abclonal, A17332, 1:1000), Cyclin D1 (Abclonal, A1301, 1:1000), Cyclin E1 (Abclonal, A14225, 1:1000), DTYMK (ZenBio, 825320, 1:1000), RRM2 (ZenBio, 382199, 1:1000), TYMS (ZenBio, 382075, 1:1000), TK1 (ZenBio, R25906, 1:1000) and β-Actin (Abclonal, AC006, 1:2000).

Techniques: Infection, Blocking Assay, Expressing

Journal: Journal of Clinical Immunology

Article Title: A Novel AGR2 Variant Causing Aberrant Monomer-Dimer Equilibrium Leading to Severe Respiratory and Digestive Symptoms

doi: 10.1007/s10875-024-01847-x

Figure Lengend Snippet: Clinical features and genetic analysis of the patients. A : Pedigree tree. B : Endoscopic imaging of Patient 1 at 18 years. Mucosa is erythematous with erosions in the lower rectum and part of the anal canal (lower panel) whereas the other intestine appeared normal (upper: ileum). C : Colon histology of Patient 1 at 15 years. Crypt distortion (crypt shortening and crypt branching), cryptitis, patchy lymphoplasmacytic inflammation and decreased goblet cells were observed. D : Chromatogram of the AGR2 variant (c.250A>C, p.(Ser84Arg)). Ref: reference, Var: variant

Article Snippet: Immunoblotting was performed with rabbit anti-AGR2 antibody (1:1000, Cell Signaling, Danvers, MA, USA), mouse anti-beta action (1:10,000, Abcam, Cambridge, UK), rabbit anti-BiP (1:5000, Cell Signaling), and horseradish peroxidase-conjugated anti-mouse or anti-rabbit IgG antibody (1:10,000; Jackson ImmunoResearch Laboratories, West Grove, PA, USA), and were detected and quantified with SuperSignal West Dura (Thermo Fisher Scientific) and a ChemiDoc Touch imaging system (BioRad, Hercules, CA, USA).

Techniques: Imaging, Variant Assay

Journal: Journal of Clinical Immunology

Article Title: A Novel AGR2 Variant Causing Aberrant Monomer-Dimer Equilibrium Leading to Severe Respiratory and Digestive Symptoms

doi: 10.1007/s10875-024-01847-x

Figure Lengend Snippet: Clinical summary of patients with biallelic AGR2 variants

Article Snippet: Immunoblotting was performed with rabbit anti-AGR2 antibody (1:1000, Cell Signaling, Danvers, MA, USA), mouse anti-beta action (1:10,000, Abcam, Cambridge, UK), rabbit anti-BiP (1:5000, Cell Signaling), and horseradish peroxidase-conjugated anti-mouse or anti-rabbit IgG antibody (1:10,000; Jackson ImmunoResearch Laboratories, West Grove, PA, USA), and were detected and quantified with SuperSignal West Dura (Thermo Fisher Scientific) and a ChemiDoc Touch imaging system (BioRad, Hercules, CA, USA).

Techniques: Variant Assay, Functional Assay, Infection, Reflux

Journal: Journal of Clinical Immunology

Article Title: A Novel AGR2 Variant Causing Aberrant Monomer-Dimer Equilibrium Leading to Severe Respiratory and Digestive Symptoms

doi: 10.1007/s10875-024-01847-x

Figure Lengend Snippet: Functional domains and motifs of AGR2 and pathogenic variants. A : AGR2 contains a cleavable signal peptide (1–20 a.a., yellow-green box), adhesion domain (21–44 a.a, sky-blue box), dimerization domain (60–64 a.a., orange box), CXXS motif (81–84 a.a., pink box), peptide binding loop (104–111 a.a., gray box), and KTEL ER retention signal (172–175 a.a., yellow box). Previously reported variants and that found in our patients are shown in black and red, respectively. B : Evolutionary conservation of the amino acid substitution

Article Snippet: Immunoblotting was performed with rabbit anti-AGR2 antibody (1:1000, Cell Signaling, Danvers, MA, USA), mouse anti-beta action (1:10,000, Abcam, Cambridge, UK), rabbit anti-BiP (1:5000, Cell Signaling), and horseradish peroxidase-conjugated anti-mouse or anti-rabbit IgG antibody (1:10,000; Jackson ImmunoResearch Laboratories, West Grove, PA, USA), and were detected and quantified with SuperSignal West Dura (Thermo Fisher Scientific) and a ChemiDoc Touch imaging system (BioRad, Hercules, CA, USA).

Techniques: Functional Assay, Binding Assay

Journal: Journal of Clinical Immunology

Article Title: A Novel AGR2 Variant Causing Aberrant Monomer-Dimer Equilibrium Leading to Severe Respiratory and Digestive Symptoms

doi: 10.1007/s10875-024-01847-x

Figure Lengend Snippet: Impact of the AGR2 Ser84Arg substitution. A - B : Whole structure and magnified view of AGR2 41 − 175 (PDB ID: 2LNS). C : Immunoblot of V5-AGR2 transiently overexpressed in HEK293T cells under reducing and non-reducing conditions. Arrows indicate the dimer of cleaved AGR2 (dC-AGR2:36 kDa), uncleaved monomer (UC-AGR2: 21 kDa), and cleaved monomer (C-AGR2:18 kDa). Representative images are shown from three independent experiments. D : Relative expression level of V5-AGR2 under reducing conditions. E : Relative expression level of V5-AGR2 dimers under non-reducing conditions. Wild-type-cleaved monomer/β-actin = 1 ( D , E ). The mean and standard deviation of three independent experiments is shown. ** P < 0.01 and **** P < 0.0001 analyzed by Dunnett’s multiple comparisons test ( D ) or ratio unpaired t-test ( E )

Article Snippet: Immunoblotting was performed with rabbit anti-AGR2 antibody (1:1000, Cell Signaling, Danvers, MA, USA), mouse anti-beta action (1:10,000, Abcam, Cambridge, UK), rabbit anti-BiP (1:5000, Cell Signaling), and horseradish peroxidase-conjugated anti-mouse or anti-rabbit IgG antibody (1:10,000; Jackson ImmunoResearch Laboratories, West Grove, PA, USA), and were detected and quantified with SuperSignal West Dura (Thermo Fisher Scientific) and a ChemiDoc Touch imaging system (BioRad, Hercules, CA, USA).

Techniques: Western Blot, Expressing, Standard Deviation

Journal: Journal of Clinical Immunology

Article Title: A Novel AGR2 Variant Causing Aberrant Monomer-Dimer Equilibrium Leading to Severe Respiratory and Digestive Symptoms

doi: 10.1007/s10875-024-01847-x

Figure Lengend Snippet: Size exclusion chromatography analysis of AGR2 variants. A : Size exclusion chromatography (SEC) profiles of AGR2 41 − 175 WT, Glu60Ala, and Ser84Arg eluted from a Superdex 200 10/300 column under reducing or non-reducing conditions. B : SEC profiles of AGR2 41 − 175 WT, Cys81Ala, and Ser84Arg under reducing or non-reducing conditions. C : Eluted fractions were analyzed using SDS-PAGE with Coomassie Brilliant Blue staining. A - C : Representative data from two independent experiments are shown

Article Snippet: Immunoblotting was performed with rabbit anti-AGR2 antibody (1:1000, Cell Signaling, Danvers, MA, USA), mouse anti-beta action (1:10,000, Abcam, Cambridge, UK), rabbit anti-BiP (1:5000, Cell Signaling), and horseradish peroxidase-conjugated anti-mouse or anti-rabbit IgG antibody (1:10,000; Jackson ImmunoResearch Laboratories, West Grove, PA, USA), and were detected and quantified with SuperSignal West Dura (Thermo Fisher Scientific) and a ChemiDoc Touch imaging system (BioRad, Hercules, CA, USA).

Techniques: Size-exclusion Chromatography, SDS Page, Staining

Journal: Journal of Clinical Immunology

Article Title: A Novel AGR2 Variant Causing Aberrant Monomer-Dimer Equilibrium Leading to Severe Respiratory and Digestive Symptoms

doi: 10.1007/s10875-024-01847-x

Figure Lengend Snippet: Mutational effects of AGR2 on ER stress and protein stability. A : Immunoblot of V5-AGR2 transiently overexpressed in HEK293T cells under reducing conditions. Arrows indicate BiP (78 kDa), UC-AGR2 (21 kDa), C-AGR2 (18 kDa), and β-actin (40 kDa) from the top. Representative images are shown from four independent experiments. B : Immunoblot of V5-AGR2 transiently overexpressed in HEK293T cells under reducing and non-reducing conditions with treatment of DMSO only or MG123. Arrows indicate dC-AGR2 (36 kDa), UC-AGR2 (21 kDa), and C-AGR2 (18 kDa) from the top. Representative images are shown from four independent experiments

Article Snippet: Immunoblotting was performed with rabbit anti-AGR2 antibody (1:1000, Cell Signaling, Danvers, MA, USA), mouse anti-beta action (1:10,000, Abcam, Cambridge, UK), rabbit anti-BiP (1:5000, Cell Signaling), and horseradish peroxidase-conjugated anti-mouse or anti-rabbit IgG antibody (1:10,000; Jackson ImmunoResearch Laboratories, West Grove, PA, USA), and were detected and quantified with SuperSignal West Dura (Thermo Fisher Scientific) and a ChemiDoc Touch imaging system (BioRad, Hercules, CA, USA).

Techniques: Western Blot

Journal: Journal of Clinical Immunology

Article Title: A Novel AGR2 Variant Causing Aberrant Monomer-Dimer Equilibrium Leading to Severe Respiratory and Digestive Symptoms

doi: 10.1007/s10875-024-01847-x

Figure Lengend Snippet: Possible pathological mechanism caused by the Ser48Arg variant. A : AGR2 monomer/dimer status of AGR wild-type and three variants (Glu60Ala, Cys81Ala, and Ser84Arg) under reducing and non-reducing conditions. B , C : Schema of mucin synthesis (upper panel) and condition of the mucosa (lower panel) in wild-type ( B ) and Ser84Arg ( C ). Mucin is multimerized through disulfide bonds between cysteine residues of mucin molecules in its normal state, where it is secreted from goblet cells into the lumen of the respiratory and digestive tracts. However, Ser84Arg AGR2 cannot catalyze this reaction and cannot produce functionally normal mucus, leading to cell damage. Blue thick line: mucin; gray triangle: abnormal dimerization domain with Glu60Ala; green ellipse: pathogens; light blue circle: AGR2; light brown square with dark brown circle: normal mucosa cell; light gray square with dark brown circle: damaged goblet cell; orange box: CXXS motif; SH: sulfate; S = S: disulfide bond; yellow square with dark brown circle: normal goblet cell; white box: dimerization domain through salt bridges (Glu60-Lys64); white dotted lines: secretion normally multimerized mucin

Article Snippet: Immunoblotting was performed with rabbit anti-AGR2 antibody (1:1000, Cell Signaling, Danvers, MA, USA), mouse anti-beta action (1:10,000, Abcam, Cambridge, UK), rabbit anti-BiP (1:5000, Cell Signaling), and horseradish peroxidase-conjugated anti-mouse or anti-rabbit IgG antibody (1:10,000; Jackson ImmunoResearch Laboratories, West Grove, PA, USA), and were detected and quantified with SuperSignal West Dura (Thermo Fisher Scientific) and a ChemiDoc Touch imaging system (BioRad, Hercules, CA, USA).

Techniques: Variant Assay