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FIX&PERM Cell Fixation and Permeabilization Kit

제품 소개

유세포분석 (flow cytometry)에서 항체를 사용한 단백질 biomarker의 세포내 염색은 연구나 진단 분야 모두에서 세포의 표현형 이나 기능을 파악하는데 새로운 옵션을 제공해 주는 방법입니다.

특정 세포 유형을 확실하게 식별하기 위해서는 세포 표면 및 세포내 마커 모두에 표지를 해야합니다.
실제로 FIX&PERM kit는 다양한 생물학적 검체 (혈액, 골수 등)에서 추출된 정상 및 악성 백혈구 집단 (leukocyte populations)의 분석에 적합합니다.

이처럼 많은 세포의 생물학적 및 기능적 연구를 위해 intracellular immunostaining이 필요합니다.
이를 위해 Nordic-MUbio/Exalpha는 동급 최고의 Cell Fixation and Permeabilization Kit를 제공합니다.

이 키트에는 Fixation Medium(시약 A) 및 Permeabilization Medium(시약 B)의 2가지 시약이 포함되어 있습니다.
먼저 시약 A로 현탁된 세포를 고정한 다음 시약 B로 세포막을 투과 시키게 됩니다.

이는 항체가 세포 내 구조에 접근할 수 있도록 하고 세포의 형태학적 산란 특성을 그대로 유지하게 합니다.

이 과정에서 특정 조성으로 만들어진 키트는 background를 줄이고 투과성 배지와 형광 색소 표지 항체를 동시에 추가할 수 있어,
cytoplasmic or nuclear enzymes, oncoproteins, cytokines, immunoglobulin 등과 같은 세포내 구조를 염색할 수 있습니다.

제품 특징

• flow cytometric scatter characteristics를 보존하면서 세포를 부드럽게 고정.
• intracellular and cell surface markers를 동시에 characterization.
• 모든 과정을 1시간 이내에 수행할 수 있으며, 즉시 분석하거나 24시간 동안 저장 가능
• 엄격한 QC 절차로 신뢰할 수 있는 결과 보장
• 최적화된 프로토콜을 가진 다양한 intracellular antibodies에 사용

• Storage: FIX&PERM® Cell Fixation and Permeabilization Kit reagents should be stored and used at room temperature. Do not freeze.
• Caution: This product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals.

• BRAND: Nordic, Exalpha, Gallus, MUbio, Biologo, ADG, ELTI, Scicons, Bii

Fixation, permeabilization, and staining procedure:

For each sample to be analyzed using 50 µl of whole blood, bone marrow, or mononuclear cell suspension in a 5 ml tube

1. Add 100 µl of Reagent A (Fixation Medium, stored and used at room temperature)
2. Incubate for 15 minutes at room temperature
3. Add 5 ml phosphate-buffered saline and centrifuge cells for 5 minutes at 300 g
4. Remove supernatant and add to cell pellet 100 µl Reagent B and 20 µl of the appropriate monoclonal antibody\
5. Vortex at low speed for 1-2 seconds
6. Incubate for 15 minutes at room temperature
7. Wash cells with PBS as above
8. Remove supernatant and resuspend cells in sheath fluid for immediate
Analysis
9. Alternatively, resuspend cells in 0.5 ml 1.0 % formaldehyde and store them at 2-8°C in the dark. Analyze cells within 24 hours

Figure 1: Flow cytometric analysis of normal bone marrow (BM) after fixation and permeabilization using GAS-002, followed by immunostaining for lactoferrin and MPO-C2.

Figure 2: Flow cytometric analysis of bone marrow (BM) from a leukemia patient after fixation and pemeabilization using GAS-002, followed by immunostaining for lactoferrin and MPO-C2.

 Figure 3: Immunostaining with Nordic-MUbio CD22-PE conjugate of undifferentiated leukemia cells of B-ALL type.

Figure 4: Immunostaining with Nordic-MUbio CD3-PE conjugate of surface CD3-negative undifferentiated leukemia cells of T-ALL type.

Figure 5: Immunostaining with Nordic-MUbio anti-MPO-FITC conjugate of undifferentiated leukemia cells of AML type.

Figure 6: Forward and side scatter pattern of a normal blood sample treated with GAS-002.

 Figure 7: Double labeling of a normal blood sample treated with GAS-002, and immunostained for Lactoferrin (PE) and MPO-C2 (FITC).

Figure 8; Double labeling of a normal blood sample treated with GAS-002, and immunostained for CD3 (PE) and MPO-C2 (FITC).

Monoclonal antibodies optimized for use with FIX&PERM

Antigen Clone Isotype Format Quantity Cat. No.
CD3 UCHT1 IgG1 Purified 0.2 mg GM-4011
FITC-labeled 100 tests GM-4012
PE-labeled 100 tests GM-4013
CD22 RFB4 IgG1 Purified 0.2 mg GM-4051
FITC-labeled 100 tests GM-4052
PE-labeled 100 tests GM-4053
Lactoferrin 4C5 IgG1 Purified 0.2 mg GM-4111
PE-labeled 100 tests GM-4113
Myeloperoxidase
(MPO-C2)
8E6 IgG1 Purified 0.2 mg GM-4191
FITC-labeled 100 tests GM-4192
PE-labeled 100 tests GM-4193
Lysozyme LZ-2 IgG1 Purified 0.2 mg GM-4131
FITC-labeled 100 tests GM-4132
CD68 Ki-M7 IgG1 FITC-labeled 100 tests GM-4152
IL-1 beta FIB3 IgG1 FITC-labeled 100 tests GM-4162
IFN-gamma GZ4 IgG1 FITC-labeled 100 tests GM-4172


Furthermore, a number of Combi panels are offered for dual immunostaining protocols.

Antigen Clone Isotype Format Quantity Cat. No.
Negative control Vl-AP IgG1 FITC-labeled 50 tests GIC-201
Vl-AP IgG1 PE-labeled
Lysozyme LZ-2 IgG1 FITC-labeled 50 tests GIC-206
Lactoferrin 4C5 IgG1 PE-labeled
Myeloperoxidase 8E6 IgG1 FITC-labeled 50 tests GIC-212
Lactoferrin 4C5 IgG1 PE-labeled
Myeloperoxidase 8E6 IgG1 FITC-labeled 50 tests GIC-213
CD3 UCHT1 IgG1 PE-labeled
Myeloperoxidase 8E6 IgG1 FITC-labeled 50 tests GIC-214
CD22 RFB4 IgG1 PE-labeled

References

Recent application paper:

Nies KPH, Kraaijvanger R, Lindelauf KHK, Drent RJMR, Rutten RMJ, Ramaekers FCS, Leers MPG. Determination of the proliferative fractions in differentiating hematopoietic cell lineages of normal bone marrow. Cytometry A. (2018) 93, 1097-1105 . doi: 10.1002/cyto.a.23564.

Mestrum SGC, de Wit NCJ, Drent RJM, Hopman AHN, Ramaekers FCS, Leers MPG. Proliferative activity is disturbed in myeloproliferative neoplasms (MPN), myelodysplastic syndrome (MDS), and MDS/MPN diseases. Differences between MDS and MDS/MPN.
Cytometry B Clin Cytom. 2020 Aug 28. doi: 10.1002/cyto.b.21946. Online ahead of print.

Further reading:

– Gerna, G., Percivalle, E., Lilleri, D., Lozza, L., Fornara, C., Hahn, G., Baldanti, F. & Revello, M. G. (2005) J Gen Virol 86, 275-84.
– Groeneveld, K., te Marvelde, J. G., van den Beemd, M. W., Hooijkaas, H. & van Dongen, J. J. (1996) Leukemia 10, 1383-9.
– Haranaga, S., Yamaguchi, H., Friedman, H., Izumi, S., & Yamamoto, Y. (2001) Infect Immun 69, 7753-9.
– Hegazy, A. N. & Klein, C. (2008) Leukemia 22, 2070-9.
– Kappelmayer, J., Gratama, J. W., Karaszi, E., Menendez, P., Ciudad, J., Rivas, R. & Orfao, A. (2000) J Immunol Methods 242, 53-65.
– Kline, M. P., Rajkumar, S. V., Timm, M. M., Kimlinger, T. K., Haug, J. L., Lust, J. A., Greipp, P. R. & Kumar, S. (2007) Leukemia 21, 1549-60
– Knapp, W., Majdic, O. & Strobl, H. (1993) Recent Results Cancer Res 131, 31-40.
– Knapp, W., Strobl, H. & Majdic, O. (1994) Cytometry 18, 187-98.
– Knapp, W., Strobl, H., Scheinecker, C., Bello-Fernandez, C. & Majdic, O. (1995) Ann Hematol 70, 281-96.
– Konikova, E., Glasova, M., Kusenda, J. & Babusikova, O. (1998) Neoplasma 45, 282-91.
– Lanza, F., Latorraca, A., Moretti, S., Castagnari, B., Ferrari, L. & Castoldi, G. (1997) Cytometry 30, 134-44.
– Millard, I., Degrave, E., Philippe, M. & Gala, J. L. (1998) Clin Chem 44, 2320-30.
– Nakase, K., Sartor, M. & Bradstock (1998) Cytometry 34, 198-202.
– Pascale, F., Contreras, V., Bonneau, M., Courbet, A., Chilmonczyk, S., Bevilacqua, C., Epardaud, M., Niborski, V., Riffault, S., Balazuc, A. M., Foulon, E., Guzylack-Piriou, L., Riteau, B., Hope, J., Bertho, N., Charley, B. & Schwartz-Cornil, I. (2008) J Immunol 180, 5963-72
– Pickl, W. F., Majdic, O., Kohl, P., Stockl, J., Riedl, E., Scheinecker, C., Bello-Fernandez, C. & Knapp, W. (1996) J Immunol 157, 3850-9.
– Riera-Sans, L., & Behrens, A. (2007) J Immunol 178, 5690-700
– Roberts, J. L., Lengi, A., Brown, S. M., Chen, M., Zhou, Y. J., O’Shea, J. J. & Buckley, R. H. (2004) Blood 103, 2009-18
– Sargent, R. L., Craig, F. E. & Swerdlow, S. H. (2009) Int J Clin Exp Pathol 2, 574-82
– Scheinecker, C., Strobl, H., Fritsch, G., Csmarits, B., Krieger, O., Majdic, O. & Knapp, W. (1995) Blood 86, 4115-23.
– Sedlmayr, P., Grosshaupt, B. & Muntean, W. (1996) Cytometry 23, 284-9.
– Strobl, H. & Knapp, W. (2004) J Biol Regul Homeost Agents 18, 335-9.
– Strobl, H., Scheinecker, C., Csmarits, B., Majdic, O. & Knapp, W. (1995) Br J Haematol 90, 774-82.
– Strobl, H., Scheinecker, C., Riedl, E., Csmarits, B., Bello-Fernandez, C., Pickl, W. F., Majdic, O. & Knapp, W. (1998) J Immunol 161, 740-8.
– Strobl, H., Takimoto, M., Majdic, O., Fritsch, G., Scheinecker, C., Hocker, P. & Knapp, W. (1993) Blood 82, 2069-78.
– Wang, X., Chang, X., Facchinetti, V., Zhuang, Y. & Su, B. (2009) J Immunol 182, 3597-608
-Fan J, Tang X, Wang Q, Zhang Z, Wu S, Li W, Liu S, Yao G, Chen H and Sun L, Mesenchymal stem cells alleviate experimental autoimmune cholangitis through immunosuppression and cytoprotective function mediated by galectin-9, Stem Cell Research & Therapy 2018, https://doi.org/10.1186/s13287-018-0979-x

Bingyu Shi, Jingjing Qi, Genhong Yao, Ruihai Feng, Zhuoya Zhang, Dandan Wang, Chen Chen, Xiaojun Tang, Liwei Lu, Wanjun Chen and Lingyun Sun
Mesenchymal stem cell transplantation ameliorates Sjögren’s syndrome via suppressing IL-12 production by dendritic cells.
Stem Cell Research & Therapy, 2018 https://doi.org/10.1186/s13287-018-1023

Rossella Zanin, Silvia Pegoraro, Gloria Ros, Yari Ciani, Silvano Piazza, Fleur Bossi, Roberta Bulla, Cristina Zennaro, Federica Tonon, Dejan Lazarevic, Elia Stupka, Riccardo Sgarra & Guidalberto Manfioletti HMGA1 promotes breast cancer angiogenesis supporting the stability, nuclear localization and transcriptional activity of FOXM1. Journal of Experimental & Clinical Cancer Research volume 38, Article number: 313 (2019)
https://doi.org/10.1186/s13046-019-1307-8

Manabu Itoh, Yosuke Mukae, Takahiro Kitsuka, Kenichi Arai, Anna Nakamura, Kazuyoshi Uchihashi, Shuji Toda, Kumika Matsubayashi, Jun-ichi Oyama, Koichi Node, Daisuke Kami, Satoshi Gojo, Shigeki Morita, Takahiro Nishida, Koichi Nakayama & Eiji Kobayashi, Development of an immunodeficient pig model allowing long-term accommodation of artificial human vascular tubes. Nature Communications volume 10, Article number: 2244 (2019) https://doi.org/10.1038/s41467-019-10107-1

CE Marked bestselling FIX&PERM ® Kits and Antibodies

All Nordic-MUbio products are manufactured according to ISO 9001 quality guidelines

Cat.No. Monoclonal Antibodies Clone
GAS-003 NM LYSE: Flow Cytometry Lysing Solution NA
GAS-003-1 NM LYSE: Flow Cytometry Lysing Solution NA
GM-4011 CD3 Mouse Ig1 single surface and single IC ab/conjugates purified UCHT1
GM-4012 CD3 Mouse Ig1 single surface and single IC ab/conjugates FITC UCHT1
GM-4013 CD3 Mouse Ig1 single surface and single IC ab/conjugates PE UCHT1
GM-4051 CD22 Mouse Ig1 single surface and single IC ab/conjugates Purified RFB4
GM-4052 CD22 Mouse Ig1 single surface and single IC ab/conjugates FITC RFB4
GM-4053 CD22 Mouse Ig1 single surface and single IC ab/conjugates PE RFB4
GM-4061 CD45 Mouse Ig2a single surface ab/conjugates Purified VIT200
GM-4062 CD45 Mouse Ig2a single surface ab/conjugates FITC VIT200
GM-4091 CD14 Mouse Ig1 single surface ab/conjugates purified MEM18
GM-4092 CD14 Mouse Ig1 single surface ab/conjugates FITC MEM18
GM-4093 CD14 Mouse Ig1 single surface ab/conjugates PE MEM18
GM-4101 CD65s Mouse IgM single surface ab/conjugates Purified VIM2
GM-4102 CD65s Mouse IgM single surface ab/conjugates FITC VIM2
GM-4172 IFN-gamma Mouse IgG1 single IC ab/conjugate FITC GZ4
GM-4152 Mouse anti CD68 (Macrosialin), conjugated to FITC Ki-M7
GM-4191 Mouse anti Myeloperoxidase (MPO) 8.00E+06
GM-4192 Mouse anti Myeloperoxidase (MPO), conjugated to FITC 8.00E+06
GM-4193 Mouse anti Myeloperoxidase (MPO), conjugated to PE 8.00E+06
GM-4131 Mouse anti Lysozyme LZ-2
GM-4132 Mouse anti-Lysozyme, conjugated to FITC LZ-2
GM-4133 Mouse anti-Lysozyme, conjugated to PE LZ-2
GM-4111 Mouse anti Lactoferrin 4C5
GM-4113 Mouse anti-Lactoferrin, conjugated to PE 4C5
Cat.No. Monoclonal Antibodies Clone
GM-4982 Mouse Ig2a Negative Control 4H1-A7
GM-4983 Mouse Ig2a Negative Control 4H1-A7
GM-4991 Mouse IgG1 Negative Control VI-AP
GM-4992 Mouse IgG1 Negative Control, conjugated to FITC VI-AP
GM-4993 Mouse IgG1 Negative Control, conjugated to PE VI-AP
GIC-201 COMBI IC Reagent: IgG1 Negative Control (FITC) and IgG1 Negative Control (PE) VI-AP/VI-AP
GIC-212 COMBI IC Reagent: Mouse anti Myeloperoxidase-C2 (FITC) and Mouse anti Lactoferrin (PE) 8E6/4C5
GIC-213 COMBI IC Reagent: Mouse anti Myeloperoxidase-C2 (FITC) and Mouse anti CD3 (PE) 8E6/UCHT1
GIC-214 COMBI IC Reagent: Mouse anti Myeloperoxidase-C2 (FITC) and Mouse anti CD22 (PE) 8E6/RFB4
GIC-206 COMBI IC Reagent: Mouse anti Lysozyme (FITC) and Mouse anti Lactoferrin (PE) LZ-2/4C5
GCT-201 COMBI Gare Control CD 45 Mouse, CD 14 Mouse VIT200/MEM18
GCT-202 COMBI IC Reagent: IgG2a Negative Control (FITC) and IgG1 Negative Control (PE) 4H1-A7/VI-AP
MUB1900P Mouse anti Vimentin RV202
MUB1900L1 Mouse anti Vimentin:FITC RV202
MUB1901P Mouse anti Vimentin RV203
MUB1902P Mouse anti Vimentin V9
MUB0311P Mouse anti alphaT- Catenin 892_24D2S
MUB0110P Mouse anti actin beta-cytoplasmic 4C2
MUB0111P Mouse anti actin gamma-cytoplasmic 2A3
MUB0329P Mouse anti Cytokeratin 19 / Keratin K19 RCK108
MUB0326P Mouse anti Cytokeratin 18 / Keratin K18 RGE53
MUB0327P Mouse anti Cytokeratin 18 / Keratin K18 RCK106
MUB0328P Mouse anti Cytokeratin 18 / Keratin K18 DE-K18
MUB0315P Mouse anti Cytokeratin 7 / Keratin K7 RCK105
MUB0316P Mouse anti Cytokeratin 7 / Keratin K7 OVTL12/30
MUB0320P Mouse anti Cytokeratin 10 / Keratin K10 DE-K10
MUB0321P Mouse anti Cytokeratin 10+13 / Keratin K10+K13 DE-K13
MUB0319P Mouse anti Cytokeratin 10 / Keratin K10 RKSE60
MUB1101P Mouse anti Lamin A 133A2
MUB1102P Mouse anti Lamin A and C 131C3
MUB1103P Mouse anti Lamin B1 119D5-F1
MUB1104P Mouse anti Lamin B2 LN43
MUB1320P Mouse anti Neu-Oncogen (C-erb B2) 9G6
MUB1500P Mouse anti P53 Bp53.12
MUB0312P Mouse anti Basal cell Cytokeratin RCK103
MUB0314P Mouse anti Cytokeratin 5 + 8 / Keratin K5 + K8 RCK102
MUB0317P Mouse anti Cytokeratin 8 / Keratin K8 H1
MUB0318P Mouse anti Cytokeratin 8 / Keratin K8 M20
MUB0323P Mouse anti Cytokeratin 14 / Keratin K14 RCK107
MUB0325P Mouse anti Cytokeratin 17 / Keratin K17 E3
MUB1310P Mouse anti Reticulon-1A / NSP-A MON160
MUB1311P Mouse anti Reticulon-1A / NSP-A MON161
MUB1313P Mouse anti Reticulon-1A/B / NSP-A/B RNL-2
MUB1314P Mouse anti Reticulon-1A/B / NSP-A/B RNL-3
MUB1315P Mouse anti Reticulon-1C / NSP-C RNL-4

주문정보

Product Code Product Name Product Type
GAS-002 FIX&PERM® Cell Fixation and Permeabilization Kit (CE) Buffers and Reagents
GM-4192 Mouse anti-Myeloperoxidase-C2 (MPO-C2), conjugated to FITC Primary Antibody
GAS-003 NM-LYSE: Flow Cytometry Lysing Solution Buffers and Reagents
GIC-212 COMBI IC Reagent: Mouse anti-Myeloperoxidase-C2 (FITC) and Mouse anti Lactoferrin (PE) Primary Antibody
GM-4132 Mouse anti Lysozyme, conjugated to FITC Primary Antibody
GM-4113 Mouse anti-Lactoferrin, conjugated to PE Primary Antibody

*상세한 제품 목록은 이곳 을 참고 해 주십시오

관련정보

제품 자료 FIXampPERM Cell Fixation and Permeabilization Kit.pdf Upload date: 2022.01.06
제품 MSDS MSDS-GAS-002.pdf Upload date: 2022.01.06