mouse monoclonal anti human cd82  (Proteintech)

Journal: Oncology Letters

Article Title: Effect and prognostic significance of the KAI1 gene in human gastric carcinoma

doi: 10.3892/ol.2015.3604

Figure Lengend Snippet: Findings of the RT-sqPCR, transfection, migration and invasion assays. (A) The pEGFP-N1-KAI1 plasmid was transfected into human gastric carcinoma SGC7901 cells by liposome. KAI1 was clearly overexpressed in pEGFP-N1-KAI1 transfected cells (lane 5) compared to the control cells (lane 4). Lanes 1, 100 bp+1kb ladder; lane 2, β-actin expression in pEGFP-N1-transfected SGC7901 cells; lane 3, β-actin expression in pEGFP-N1-KAI1-transfected SGC7901 cells; lane 4, KAI1 expression in pEGFP-N1-transfected SGC7901 cells; and lane 5, KAI1 expression in pEGFP-N1-KAI1-transfected SGC7901 cells. KAI1 was clearly more highly expressed in lane 5 compared with lane 4. (B) Fluorescent expression of pEGFP-N1-KAI1 and pEGFP-N1 in transfected SGC7901 cells, revealing that transfection with the KAI1 gene inhibited the migration and invasion activity of SGC7901 cells. (C) The invasion activity of pEGFP-N1-KAI1 cells was significantly downregulated. (D) The migratory activity of pEGFP-N1-KAI1 cells was significantly decreased. RT-sqPCR, reverse transcription-semi-quantitative polymerase chain reaction; KAI1, Kangai 1.

Article Snippet: The monoclonal mouse anti-human KAI1/CD82 antibody (cat no. 564341; BD Biosciences, San Jose, CA, USA) was diluted at 1:200.

Techniques: Transfection, Migration, Plasmid Preparation, Expressing, Activity Assay, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Effect and prognostic significance of the KAI1 gene in human gastric carcinoma

doi: 10.3892/ol.2015.3604

Figure Lengend Snippet: Association between KAI1 expression and the differentiation of gastric cancer tumors.

Article Snippet: The monoclonal mouse anti-human KAI1/CD82 antibody (cat no. 564341; BD Biosciences, San Jose, CA, USA) was diluted at 1:200.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Effect and prognostic significance of the KAI1 gene in human gastric carcinoma

doi: 10.3892/ol.2015.3604

Figure Lengend Snippet: Association between KAI1 expression and other clinicopathological features of gastric cancer.

Article Snippet: The monoclonal mouse anti-human KAI1/CD82 antibody (cat no. 564341; BD Biosciences, San Jose, CA, USA) was diluted at 1:200.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Effect and prognostic significance of the KAI1 gene in human gastric carcinoma

doi: 10.3892/ol.2015.3604

Figure Lengend Snippet: KAI1 mRNA expression in gastric cancer tissue and its correlation with clinical pathology.

Article Snippet: The monoclonal mouse anti-human KAI1/CD82 antibody (cat no. 564341; BD Biosciences, San Jose, CA, USA) was diluted at 1:200.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Effect and prognostic significance of the KAI1 gene in human gastric carcinoma

doi: 10.3892/ol.2015.3604

Figure Lengend Snippet: KAI1 protein and mRNA expression in gastric cancer tissue and the correlation with the prognosis of gastric cancer patients. (A) Positive staining for the expression of the KAI1 protein was brown in color and diffusely distributed, with staining being located in the cytoplasm and plasma membrane (SP 400). (B) KAI1 mRNA positive staining was mainly located in cytoplasm, and cells expressing KAI1 were brown granular with clear location. (C) Effect of KAI1 protein expression on the survival time in gastric cancer patients. In total, 28 gastric cancer patients demonstrated KAI1 expression and 100 gastric cancer patients did not express KAI1. The survival time of the group that expressed the KAI1 protein was significantly longer compared with the group that did not express the KAI1 protein (P<0.05). (D) Effect of KAI1 mRNA expression on the survival time in gastric cancer. In total, 40 gastric cancer patients expressed KAI1 mRNA and 88 gastric cancer patients did not express KAI1 mRNA. The survival time of the group that expressed KAI1 mRNA was significantly longer compared with the group that did not express KAI1 mRNA (P<0.05). KAI1, Kangai 1.

Article Snippet: The monoclonal mouse anti-human KAI1/CD82 antibody (cat no. 564341; BD Biosciences, San Jose, CA, USA) was diluted at 1:200.

Techniques: Expressing, Staining

Journal: Oncology Letters

Article Title: Effect and prognostic significance of the KAI1 gene in human gastric carcinoma

doi: 10.3892/ol.2015.3604

Figure Lengend Snippet: Association between KAI1 expression and the survival time of patients.

Article Snippet: The monoclonal mouse anti-human KAI1/CD82 antibody (cat no. 564341; BD Biosciences, San Jose, CA, USA) was diluted at 1:200.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Effect and prognostic significance of the KAI1 gene in human gastric carcinoma

doi: 10.3892/ol.2015.3604

Figure Lengend Snippet: The association between KAI1 mRNA expression and the survival time of patients.

Article Snippet: The monoclonal mouse anti-human KAI1/CD82 antibody (cat no. 564341; BD Biosciences, San Jose, CA, USA) was diluted at 1:200.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Effect and prognostic significance of the KAI1 gene in human gastric carcinoma

doi: 10.3892/ol.2015.3604

Figure Lengend Snippet: Effect of KAl1 gene transfection on the expression of genes associated with gastric cancer metastasis.

Article Snippet: The monoclonal mouse anti-human KAI1/CD82 antibody (cat no. 564341; BD Biosciences, San Jose, CA, USA) was diluted at 1:200.

Techniques: Transfection, Expressing

Journal: Oncology Letters

Article Title: Clinical significance of KAI1/CD82 protein expression in nasopharyngeal carcinoma

doi: 10.3892/ol.2015.2891

Figure Lengend Snippet: KAI1/CD82 protein expression in human nasopharyngeal carcinoma in (A) SUNE-1-6-10B, (B) CNE-1, (C) CNE-2Z, (D) SUNE-1 and (E) SUNE-1-5-8F cell lines with different metastatic potentials (magnification, ×100).

Article Snippet: After three washes with phosphate buffered-saline (PBS; Fuzhou Maixin Biotechnology Development Co., Ltd., Fuzhou, China) for 15 min each, l% bovine serum albumin (Fuzhou Maixin Biotechnology Development Co., Ltd.) was used to block non-specific antigen activity for 2 h. After blocking, the membranes were incubated with mouse anti-human KAI1/CD82 monoclonal primary antibody (l:250; sc-17752; Santa Cruz Biotechnology, Inc., CA, USA) overnight at 4°C, followed by washing with PBS.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Clinical significance of KAI1/CD82 protein expression in nasopharyngeal carcinoma

doi: 10.3892/ol.2015.2891

Figure Lengend Snippet: Comparison of KAI1/CD82 protein positive expression rates in five cell lines.

Article Snippet: After three washes with phosphate buffered-saline (PBS; Fuzhou Maixin Biotechnology Development Co., Ltd., Fuzhou, China) for 15 min each, l% bovine serum albumin (Fuzhou Maixin Biotechnology Development Co., Ltd.) was used to block non-specific antigen activity for 2 h. After blocking, the membranes were incubated with mouse anti-human KAI1/CD82 monoclonal primary antibody (l:250; sc-17752; Santa Cruz Biotechnology, Inc., CA, USA) overnight at 4°C, followed by washing with PBS.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Clinical significance of KAI1/CD82 protein expression in nasopharyngeal carcinoma

doi: 10.3892/ol.2015.2891

Figure Lengend Snippet: KAI1/CD82 gene expression in different human nasopharyngeal carcinoma cell lines. (A) SUNE-1-5-8F, (B) CNE-2Z, (C) SUNE-1, (D) CNE-1 and (E) SUNE-1-6-10B.

Article Snippet: After three washes with phosphate buffered-saline (PBS; Fuzhou Maixin Biotechnology Development Co., Ltd., Fuzhou, China) for 15 min each, l% bovine serum albumin (Fuzhou Maixin Biotechnology Development Co., Ltd.) was used to block non-specific antigen activity for 2 h. After blocking, the membranes were incubated with mouse anti-human KAI1/CD82 monoclonal primary antibody (l:250; sc-17752; Santa Cruz Biotechnology, Inc., CA, USA) overnight at 4°C, followed by washing with PBS.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Clinical significance of KAI1/CD82 protein expression in nasopharyngeal carcinoma

doi: 10.3892/ol.2015.2891

Figure Lengend Snippet: Expression level changes of KAI1/CD82 gene in different human nasopharyngeal carcinoma cell lines.

Article Snippet: After three washes with phosphate buffered-saline (PBS; Fuzhou Maixin Biotechnology Development Co., Ltd., Fuzhou, China) for 15 min each, l% bovine serum albumin (Fuzhou Maixin Biotechnology Development Co., Ltd.) was used to block non-specific antigen activity for 2 h. After blocking, the membranes were incubated with mouse anti-human KAI1/CD82 monoclonal primary antibody (l:250; sc-17752; Santa Cruz Biotechnology, Inc., CA, USA) overnight at 4°C, followed by washing with PBS.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Clinical significance of KAI1/CD82 protein expression in nasopharyngeal carcinoma

doi: 10.3892/ol.2015.2891

Figure Lengend Snippet: Positive KAI1/CD82 protein expression was identified in (A) non-neoplastic nasopharyngeal and (B) NPC tissues by immunohistochemistry. (magnification: A, ×40; B, ×10).

Article Snippet: After three washes with phosphate buffered-saline (PBS; Fuzhou Maixin Biotechnology Development Co., Ltd., Fuzhou, China) for 15 min each, l% bovine serum albumin (Fuzhou Maixin Biotechnology Development Co., Ltd.) was used to block non-specific antigen activity for 2 h. After blocking, the membranes were incubated with mouse anti-human KAI1/CD82 monoclonal primary antibody (l:250; sc-17752; Santa Cruz Biotechnology, Inc., CA, USA) overnight at 4°C, followed by washing with PBS.

Techniques: Expressing, Immunohistochemistry

Journal: Oncology Letters

Article Title: Clinical significance of KAI1/CD82 protein expression in nasopharyngeal carcinoma

doi: 10.3892/ol.2015.2891

Figure Lengend Snippet: Association between KAI1/CD82 protein expression and patient clinical parameters.

Article Snippet: After three washes with phosphate buffered-saline (PBS; Fuzhou Maixin Biotechnology Development Co., Ltd., Fuzhou, China) for 15 min each, l% bovine serum albumin (Fuzhou Maixin Biotechnology Development Co., Ltd.) was used to block non-specific antigen activity for 2 h. After blocking, the membranes were incubated with mouse anti-human KAI1/CD82 monoclonal primary antibody (l:250; sc-17752; Santa Cruz Biotechnology, Inc., CA, USA) overnight at 4°C, followed by washing with PBS.

Techniques: Expressing

Journal: Journal of Molecular Endocrinology

Article Title: CD82 gene suppression in endometrial stromal cells leads to increase of the cell invasiveness in the endometriotic milieu

doi: 10.1530/jme-10-0165

Figure Lengend Snippet: Figure 1 The expression of CD82 is decreased in the primary ESCs from eutopic endometrium and endometriotic tissues. CD82 mRNA and protein levels in human ESCs were analyzed by quantitative real-time PCR (a), western blot (b), or in-cell Western (c). The relative fold mRNA and protein levels of CD82 in ESCs were compared between the eutopic ESCs, ectopic ESCs, and normal ESCs. CD82 (red) and actin (green). Results were highly reproducible in three independent experiments. In this study, normal, ESCs from endometrium without endometriosis; eutopic and ectopic, ESCs from eutopic endometrium and ectopic tissue with endometriosis respectively. Error bars depict the S.E.M. *P!0.05 and **P!0.01 compared with the control. #P!0.05 and ##P!0.01 compared with the eutopic.

Article Snippet: The resolved protein was transferred onto polyvinylidene difluoride membranes (Bio-Rad), and incubated with a 1:500 dilution of mouse anti-human CD82 monoclonal antibody (SC-15572, Santa Cruz Biotechnology) and a 1:1000 dilution of mouse antihuman b-actin monoclonal antibody (Santa Cruz Biotechnology) in PBS containing 0.05% Tween-20 and 5% FCS respectively.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, In-Cell ELISA, Control

Journal: Journal of Molecular Endocrinology

Article Title: CD82 gene suppression in endometrial stromal cells leads to increase of the cell invasiveness in the endometriotic milieu

doi: 10.1530/jme-10-0165

Figure Lengend Snippet: Figure 2 The combination of TCDD with 17b-estradiol or co-culture with U937 downregulates CD82 expression in ESCs. (a–c) ESCs were treated, respectively, with different concentrations of TCDD (from 10K12 to 10K8 M), 17b-estradiol (from 10K11 to 10K7 M) or the combination of 10K9 M TCDD and 10K8 M 17b-estradiol for 48 h, with vehicle as controls. In-cell Western indicates that TCDD alone can significantly decrease, and 17b-estradiol alone can increase the expression of CD82 in ESCs, but 17b-estradiol can coordinate with TCDD in downregulating CD82 expression. (d) ESCs (1!105

Article Snippet: The resolved protein was transferred onto polyvinylidene difluoride membranes (Bio-Rad), and incubated with a 1:500 dilution of mouse anti-human CD82 monoclonal antibody (SC-15572, Santa Cruz Biotechnology) and a 1:1000 dilution of mouse antihuman b-actin monoclonal antibody (Santa Cruz Biotechnology) in PBS containing 0.05% Tween-20 and 5% FCS respectively.

Techniques: Co-Culture Assay, Expressing, In-Cell ELISA

Journal: Journal of Molecular Endocrinology

Article Title: CD82 gene suppression in endometrial stromal cells leads to increase of the cell invasiveness in the endometriotic milieu

doi: 10.1530/jme-10-0165

Figure Lengend Snippet: Figure 3 CD82 silence enhances the invasion of ESCs. The ESCs from endometrium without endometriosis were silenced for CD82 for 72 h, and the efficiency was demonstrated by in-cell Western (a) and western blot (b) respectively. CD82 (red) and actin (green). (c) After CD82 in these ESCs was silenced, the invasiveness of ESCs was enhanced apparently. (d) After CD82 had been knocked down, ESCs were treated with TCDD, 17b-estradiol or the combination of both for another 48 h, and then the invasiveness of ESCs was detected. TCDD, 17b-estradiol or the combination can promote the invasion of ESCs. Moreover, TCDD or the combination shows a synergistic effect with the CD82 silence in elevating invasiveness (the ratio of the cells migrated to the lower chamber to the protein concentration of total cells) of ESCs. These pictures are representatives of three individual experiments. Blank, no transfection; control, the non-targeting siRNA oligonucleotides; silence, CD82 is knocked down. Error bars depict the S.E.M. *P!0.05 and **P!0.01 compared with the negative control. #P!0.05 and ##P!0.01 compared with the CD82 silence.

Article Snippet: The resolved protein was transferred onto polyvinylidene difluoride membranes (Bio-Rad), and incubated with a 1:500 dilution of mouse anti-human CD82 monoclonal antibody (SC-15572, Santa Cruz Biotechnology) and a 1:1000 dilution of mouse antihuman b-actin monoclonal antibody (Santa Cruz Biotechnology) in PBS containing 0.05% Tween-20 and 5% FCS respectively.

Techniques: In-Cell ELISA, Western Blot, Protein Concentration, Transfection, Control, Negative Control

Journal: Journal of Molecular Endocrinology

Article Title: CD82 gene suppression in endometrial stromal cells leads to increase of the cell invasiveness in the endometriotic milieu

doi: 10.1530/jme-10-0165

Figure Lengend Snippet: Figure 4 CD82 suppresses ESCs invasion by inhibiting CCL2 secretion and CCR2 expression. (a and b) CCL2 secretion and CCR2 expression in ESCs from endometrium without endometriosis were increased at 120 h after the CD82 silence. The CCL2 level was calculated as the ratio of the secretion of CCL2 in supernatant by ELISA to the protein concentration of total cells. CCR2 (green) and actin (red). ESCs were treated with anti-CCL2 neutralizing antibody (0, 0.2, 1, and 5 mg/ml) or RS102895 (0, 10, 50, 100, and 500 ng/ml), then the invasion (c and d) of ESCs was detected by Matrigel invasion assay. The results indicate that both RS102895 and anti-CCL2 neutralizing antibody can inhibit the invasiveness of ESCs. (e) After CD82 was silenced in ESCs, the ESCs were treated, respectively, with anti-CCL2 neutralizing antibody at 1 mg/ml or RS102895 at 100 ng/ml for another 48 h, and then the invasiveness of ESCs was detected. RS102895 or anti-CCL2 neutralizing antibody can block the increased invasiveness of ESCs induced by CD82 silence. The cell invasiveness was calculated as the ratio of the cells migrated to the lower chamber to the protein concentration of total cells. The pictures are representatives of three individual experiments. In this study, control, the non-targeting siRNA oligonucleotides; silence, CD82 is knocked down. Error bars depict the S.E.M. *P!0.05 and **P!0.01 compared with the control. ##P!0.01 compared with CD82 silence.

Article Snippet: The resolved protein was transferred onto polyvinylidene difluoride membranes (Bio-Rad), and incubated with a 1:500 dilution of mouse anti-human CD82 monoclonal antibody (SC-15572, Santa Cruz Biotechnology) and a 1:1000 dilution of mouse antihuman b-actin monoclonal antibody (Santa Cruz Biotechnology) in PBS containing 0.05% Tween-20 and 5% FCS respectively.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Protein Concentration, Invasion Assay, Blocking Assay, Control

Journal: Journal of Molecular Endocrinology

Article Title: CD82 gene suppression in endometrial stromal cells leads to increase of the cell invasiveness in the endometriotic milieu

doi: 10.1530/jme-10-0165

Figure Lengend Snippet: Figure 5 CD82 inhibits the invasion of ESCs by downregulating CCL2 secretion and CCR2 expression via MAPK and integrinb1 signal pathway. (a) CD82 upregulates TIMP1 and TIMP2 expression, downregulates integrinb1 and integrinanb3 expression in ESCs, but has no influence on MMP2 and MMP9 expression. In this study, MMP2, MMP9, TIMP1, TIMP2, integrinb1, and integrinanb3 are shown in red and actin is shown in green. (b) After the silence, ESCs were treated with U0126 or anti-integrinb1 neutralizing antibody for another 24 h, and then the CCL2 secretion and CCR2 expression were detected by ELISA and in-cell Western, respectively, and the invasion of ESCs (d) by invasion assay. The results show that U0126 or anti-integrinb1 neutralizing antibody can abolish completely the increase of CCL2 secretion, CCR2 expression and invasion of ESCs induced by CD82 silence. CCR2 (green) and actin (red). (c) We treated CD82-silenced ESCs with U0126 or anti-integrinb1 neutralizing antibody for another 24 h, or anti-CCL2 neutralizing antibody or RS102895 for another 48h, and then TIMP1 and TIMP2 expression were evaluated by in-cell Western. TIMP1 and TIMP2 (red) and actin (green). The CCL2 level was calculated as the ratio of the secretion of CCL2 in supernatant by ELISA to the protein concentration of total cells. The cell invasiveness was calculated as the ratio of the cells migrated to the lower chamber to the protein concentration of total cells. Results were highly reproducible in three independent experiments. In this study, control, the non-targeting siRNA oligonucleotides; silence, CD82 is knocked down. Error bars depict the S.E.M. *P!0.05 and **P!0.01 compared with the negative control. ##P!0.01 compared with CD82 silence.

Article Snippet: The resolved protein was transferred onto polyvinylidene difluoride membranes (Bio-Rad), and incubated with a 1:500 dilution of mouse anti-human CD82 monoclonal antibody (SC-15572, Santa Cruz Biotechnology) and a 1:1000 dilution of mouse antihuman b-actin monoclonal antibody (Santa Cruz Biotechnology) in PBS containing 0.05% Tween-20 and 5% FCS respectively.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, In-Cell ELISA, Invasion Assay, Protein Concentration, Control, Negative Control

Journal: Journal of Molecular Endocrinology

Article Title: CD82 gene suppression in endometrial stromal cells leads to increase of the cell invasiveness in the endometriotic milieu

doi: 10.1530/jme-10-0165

Figure Lengend Snippet: Figure 6 The combination of TCDD with 17b-estradiol stimulates CCL2 secretion, CCR2 expression and invasion of ESCs by downregulating CD82 expression. (a and b) After CD82 was silenced in normal ESCs (nZ6), the ESCs were treated with TCDD, 17b-estradiol or the combination of both for another 48 h, and then CCL2 secretion and CCR2 expression of ESCs were detected by ELISA and in-cell Western respectively. TCDD not only stimulates CCL2 secretion and CCR2 expression in ESCs, but also recovers the increase of CCL2 secretion and CCR2 expression in ESCs induced by CD82 silence. CCR2 (green) and actin (red). (c) The ESCs were treated with TCDD, 17b-estradiol, or the combination of both with RS102895 or/and anti-CCL2 neutralizing antibody for 48 h respectively. Thereafter, the invasion of ESCs was detected by Matrigel invasion assay. The results indicate that either RS102895 or anti-CCL2 neutralizing antibody abolishes completely the invasion enhancement of ESCs induced by the combination of TCDD and 17b-estradiol. The CCL2 level was calculated as the ratio of the secretion of CCL2 in supernatant by ELISA to the protein concentration of total cells. Meanwhile, the cell invasiveness was calculated by the ratio of the cells migrated to the lower chamber to the protein concentration of total cells. These pictures are representatives of three individual experiments. Error bars depict the S.E.M. *P!0.05 and **P!0.01 compared with the negative control. #P!0.05 and ##P!0.01 compared with CD82 silence. DP!0.05 compared with E2 treat- ment group.

Article Snippet: The resolved protein was transferred onto polyvinylidene difluoride membranes (Bio-Rad), and incubated with a 1:500 dilution of mouse anti-human CD82 monoclonal antibody (SC-15572, Santa Cruz Biotechnology) and a 1:1000 dilution of mouse antihuman b-actin monoclonal antibody (Santa Cruz Biotechnology) in PBS containing 0.05% Tween-20 and 5% FCS respectively.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, In-Cell ELISA, Invasion Assay, Protein Concentration, Negative Control

Journal: Journal of Molecular Endocrinology

Article Title: CD82 gene suppression in endometrial stromal cells leads to increase of the cell invasiveness in the endometriotic milieu

doi: 10.1530/jme-10-0165

Figure Lengend Snippet: Figure 7 Schematic roles of CD82 regulation in the origin and development of endometriosis. The ESCs with the abnormal repressed CD82 expression owing to inherent defects or the combined effect of TCDD and estrogen regurgitates into the peritoneal cavity, which upregulates CCL2 secretion and CCR2 expression through MAPK and integrinb1 signal pathway. Downregulation of TIMP1 and TIMP2 as effective molecules is involved in the invasiveness increase of ESCs. The increased CCL2 secretion recruits more macrophages into the peritoneal cavity in a paracrine manner, and these cells further downregulate CD82 expression in ESCs, which leads to a vicious circle, and eventually the onset and development of endometriosis.

Article Snippet: The resolved protein was transferred onto polyvinylidene difluoride membranes (Bio-Rad), and incubated with a 1:500 dilution of mouse anti-human CD82 monoclonal antibody (SC-15572, Santa Cruz Biotechnology) and a 1:1000 dilution of mouse antihuman b-actin monoclonal antibody (Santa Cruz Biotechnology) in PBS containing 0.05% Tween-20 and 5% FCS respectively.

Techniques: Expressing