Mouse igg1k mopc 21

Manufactured by Abcam

Sourced in United States

Mouse IgG1k (MOPC-21) is a purified monoclonal antibody derived from the MOPC-21 mouse myeloma cell line. It is an immunoglobulin G1 kappa (IgG1k) isotype.

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Lab products found in correlation

Mouse monoclonal anti cxcr4, by Abcam (4 mentions) Rabbit polyclonal anti cxcr7, by GeneTex (3 mentions) Alexa fluor 488 anti rabbit, by Thermo Fisher Scientific (2 mentions) Rabbit immunoglobulin fraction code x0903, by Agilent Technologies (2 mentions) Axioplan 2, by Zeiss (2 mentions)

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MOPC-21

4 protocols using mouse igg1k mopc 21

CXCR4 and CXCR7 Expression in Tissue Microarrays

Cited 1 time

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All formalin-fixed paraffin-embedded tissue samples were provided from the Institute of Pathology, University Hospital Duesseldorf. The construction of the tissue microarrays, immunohistochemistry and analysis of protein expression using the immunoreactivity score (IRS) reported by Remmele (Remmele et al, 1986 (link)) were performed as described previously (Werner et al, 2016 (link)). The staining intensity of the different samples was reviewed by two independent investigators (TW and CF) in a blinded manner.
For immunohistochemical staining the following primary antibodies were used: mouse monoclonal anti-CXCR4 (1 : 100 dilution; Abcam, Cambridge, UK), rabbit polyclonal anti-CXCR7 (1 : 200 dilution; GeneTex, Irvine, CA, USA).
Isotype controls were performed using mouse IgG1k (MOPC-21; 1 : 50 dilution; Abcam) or rabbit immunoglobulin fraction (Code X0903; 1 : 1000 dilution; Dako, Glostrup, Denmark). CXCR4-expressing tonsil tissue and CXCR7-expressing pancreatic adenocarcinoma served as positive controls. The prognostic power of CXCR4 and CXCR7 was assessed according to the REporting recommendations for tumour MARKer prognostic studies (REMARK) (McShane et al, 2005 (link)).

Werner T.A., Forster C.M., Dizdar L., Verde P.E., Raba K., Schott M., Knoefel W.T, & Krieg A. (2017). CXCR4/CXCR7/CXCL12 axis promotes an invasive phenotype in medullary thyroid carcinoma. British Journal of Cancer, 117(12), 1837-1845.

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Immunofluorescence Staining of Chemokine Receptors

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Cells were fixed with ice-cold methanol and incubated with 0.5% Triton. Unspecific epitops were blocked using 20% AB serum. Next, cells were incubated with their respective primary antibody or isotype control (mouse monoclonal anti-CXCR4, 1 : 250 dilution; Abcam; rabbit polyclonal anti-CXCR7, 1 : 500 dilution; GeneTex; mouse IgG1k (MOPC-21); 2 μg ml⁻¹; Abcam; rabbit immunoglobulin fraction (Code X0903) 2 μg ml⁻¹; Dako). After 45 min of incubation, cells were washed and treated with either anti-mouse- or anti-rabbit-Alexa Fluor 488 (10 μg ml⁻¹; Thermo Fisher). Nuclear staining was achieved using DAPI (200 ng ml⁻¹). After fixation with 1% PFA, the cells were visualised using a fluorescence microscope at × 400 magnification (Zeiss Axioplan 2, Carl Zeiss).

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Immunofluorescence Analysis of CXCR4 and CXCR7

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Cells were cultivated in 6-well chamber slides and fixated with ice-cold methanol. After treatment with 0.5 % Triton the cells were blocked with 20 % AB serum and incubated with their respective primary antibody and isotype control (mouse monoclonal anti-CXCR4, 1:250 dilution; Abcam, Cambridge, UK; rabbit polyclonal anti-CXCR7, 1:500 dilution; GeneTex, Irvine, CA, USA; mouse IgG1k (MOPC-21); 2 µg/ml; Abcam, Cambridge, UK; rabbit immunoglobulin fraction (Code X0903) 2 µg/ml; Dako, Glostrup, Denmark). After 45 minutes of incubation, cells were washed and incubated with anti-mouse- or anti-rabbit-Alexa Fluor® 488 (10 µg/ml; Thermo Fisher, MA, USA), respectively. Nuclear staining was carried out with DAPI (200 ng/ml). After fixating the cells with 1 % PFA, visualization was achieved using a fluorescence microscope at 400x magnification (Zeiss Axioplan 2, Carl Zeiss, Göttingen, Germany).

Werner T.A., Forster C.M., Dizdar L., Verde P.E., Raba K., Schott M., Knoefel W.T, & Krieg A. (2018). CXCR4/CXCR7/CXCL12-Axis in Follicular Thyroid Carcinoma. Journal of Cancer, 9(6), 929-940.

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Immunohistochemical Analysis of CXCR4 and CXCR7 in Follicular Thyroid Carcinoma

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Formalin-fixed paraffin-embedded tissue samples were obtained from the Institute of Pathology, University Hospital Duesseldorf. All tissue samples were newly reviewed prior to this study and the diagnosis of FTC histologically confirmed. FTCs were defined as thyroid carcinomas with follicular differentiation in the absence of papillary nuclear features, with either capsular or vascular invasion. The construction of the tissue microarray, immunohistochemistry and protein expression analyses were performed as described previously21 (link),22 (link). The expression levels of CXCR4 and CXCR7 were scored by two independent investigators (TW, CF) using the immunoreactivity score (IRS) reported by Remmele23 (link) in a blinded manner.
The immunohistochemistry was carried out using mouse monoclonal anti-CXCR4 (1:100 dilution; Abcam, Cambridge, UK) and rabbit polyclonal anti-CXCR7 (1:200 dilution; GeneTex, Irvine, CA, USA) as primary antibodies. Isotype control was performed using mouse IgG1k (MOPC-21; 1:50 dilution; Abcam, Cambridge, UK) and rabbit immunoglobulin fraction (Code X0903; 1:1000 dilution; Dako, Glostrup, Denmark). CXCR4 expressing tonsil tissue and CXCR7 expressing pancreatic adenocarcinoma served as positive controls. The prognostic power of CXCR4 and CXCR7 was assessed in accordance with the REporting recommendations for tumour MARKer prognostic studies (REMARK)24 .

Werner T.A., Forster C.M., Dizdar L., Verde P.E., Raba K., Schott M., Knoefel W.T, & Krieg A. (2018). CXCR4/CXCR7/CXCL12-Axis in Follicular Thyroid Carcinoma. Journal of Cancer, 9(6), 929-940.

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