MA1-21350

antibody from Invitrogen Antibodies

Targeting: ZAP70 SRK, STD, ZAP-70

Provider product page for MA1-21350

Western blot

ELISA

Immunocytochemistry

Immunoprecipitation

Flow cytometry

Antibody data

Antibody Data
Antigen structure
References [34]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [1]

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Product number: MA1-21350 - Provider product page
Provider: Invitrogen Antibodies
Product name: Zap-70 Monoclonal Antibody (1E7.2)
Antibody type: Monoclonal
Antigen: Other
Description: Positive control of Jurkat cell lysate suggested.
Reactivity: Human, Mouse, Rat
Host: Mouse
Isotype: IgG
Antibody clone number: 1E7.2
Vial size: 100 µg
Concentration: 1.7 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

ZAP70 protein structure - MA1-21350 shown in red.

Submitted references Pitfalls and limitations of ZAP-70 detection in chronic lymphocytic leukemia.

Vroblova V, Smolej L, Krejsek J
Hematology (Amsterdam, Netherlands) 2012 Sep;17(5):268-74

Quantitative analysis of protein phosphorylations and interactions by multi-colour IP-FCM as an input for kinetic modelling of signalling networks.

Deswal S, Schulze AK, Höfer T, Schamel WW
PloS one 2011;6(7):e22928

Methodological comparison of two anti-ZAP-70 antibodies.

Degheidy HA, Venzon DJ, Farooqui MZ, Abbasi F, Arthur DC, Wilson WH, Wiestner A, Stetler-Stevenson MA, Marti GE
Cytometry. Part B, Clinical cytometry 2011 Sep;80(5):300-8

Prospective study of clinical and biological prognostic factors at diagnosis in patients with early stage B-cell chronic lymphocytic leukemia.

Oliveira AC, de la Banda E, Domingo-Domenech E, Encuentra M, Mercadal S, Domingo A, Alonso E, Espinet B, Grau J, De Sevilla AF, Gonzalez-Barca E
Leukemia & lymphoma 2011 Mar;52(3):429-35

High expression of the immature laminin receptor protein correlates with mutated IGVH status and predicts a favorable prognosis in chronic lymphocytic leukemia.

Friedrichs B, Siegel S, Reimer R, Barsoum A, Coggin J Jr, Kabelitz D, Heidorn K, Schulte C, Schmitz N, Zeis M
Leukemia research 2011 Jun;35(6):721-9

Cluster analysis of immunophenotypic data: the example of chronic lymphocytic leukemia.

Zucchetto A, Cattarossi I, Nanni P, Zaina E, Prato G, Gilestro M, Marconi D, Bulian P, Rossi FM, Del Vecchio L, Omedè P, Geuna M, Del Poeta G, Gattei V
Immunology letters 2011 Jan 30;134(2):137-44

Significant change in ZAP-70 expression during the course of chronic lymphocytic leukemia.

Vroblova V, Vrbacky F, Hrudkova M, Jankovicova K, Schmitzova D, Maly J, Krejsek J, Smolej L
European journal of haematology 2010 Jun;84(6):513-7

Recruitment of PKC-betaII to lipid rafts mediates apoptosis-resistance in chronic lymphocytic leukemia expressing ZAP-70.

zum Büschenfelde CM, Wagner M, Lutzny G, Oelsner M, Feuerstacke Y, Decker T, Bogner C, Peschel C, Ringshausen I
Leukemia 2010 Jan;24(1):141-52

Humoral immune responses against the immature laminin receptor protein show prognostic significance in patients with chronic lymphocytic leukemia.

Friedrichs B, Siegel S, Kloess M, Barsoum A, Coggin J Jr, Rohrer J, Jakob I, Tiemann M, Heidorn K, Schulte C, Kabelitz D, Steinmann J, Schmitz N, Zeis M
Journal of immunology (Baltimore, Md. : 1950) 2008 May 1;180(9):6374-84

Humoral immune responses against the immature laminin receptor protein show prognostic significance in patients with chronic lymphocytic leukemia.

Optimization of flow cytometric measurement of ZAP-70 in chronic lymphocytic leukemia.

Preobrazhensky SN, Bahler DW
Cytometry. Part B, Clinical cytometry 2008 Mar;74(2):118-27

FCRL2 expression predicts IGHV mutation status and clinical progression in chronic lymphocytic leukemia.

Li FJ, Ding S, Pan J, Shakhmatov MA, Kashentseva E, Wu J, Li Y, Soong SJ, Chiorazzi N, Davis RS
Blood 2008 Jul 1;112(1):179-87

An improved flow cytometric method using FACS Lysing Solution for measurement of ZAP-70 expression in B-cell chronic lymphocytic leukemia.

Bekkema R, Tadema A, Daenen SM, Kluin-Nelemans HC, Mulder AB
Cytometry. Part B, Clinical cytometry 2008 Jan;74(1):40-4

Multicenter study of ZAP-70 expression in patients with B-cell chronic lymphocytic leukemia using an optimized flow cytometry method.

Gachard N, Salviat A, Boutet C, Arnoulet C, Durrieu F, Lenormand B, Leprêtre S, Olschwang S, Jardin F, Lafage-Pochitaloff M, Penther D, Sainty D, Reminieras L, Feuillard J, Béné MC, GEIL
Haematologica 2008 Feb;93(2):215-23

Comprehensive biomarker and genomic analysis identifies p53 status as the major determinant of response to MDM2 inhibitors in chronic lymphocytic leukemia.

Saddler C, Ouillette P, Kujawski L, Shangary S, Talpaz M, Kaminski M, Erba H, Shedden K, Wang S, Malek SN
Blood 2008 Feb 1;111(3):1584-93

National standardization of ZAP-70 determination by flow cytometry: the French experience.

Le Garff-Tavernier M, Ticchioni M, Brissard M, Salmon C, Raynaud S, Davi F, Bernard A, Merle-Béral H, Ajchenbaum-Cymbalista F, Letestu R
Cytometry. Part B, Clinical cytometry 2007 Mar;72(2):103-8

ZAP-70 expression, as detected by immunohistochemistry on bone marrow biopsies from early-phase CLL patients, is a strong adverse prognostic factor.

Zanotti R, Ambrosetti A, Lestani M, Ghia P, Pattaro C, Remo A, Zanetti F, Stella S, Perbellini O, Prato G, Guida G, Caligaris-Cappio F, Menestrina F, Pizzolo G, Chilosi M
Leukemia 2007 Jan;21(1):102-9

Familial B-cell chronic lymphocytic leukemia: analysis of cytogenetic abnormalities, immunophenotypic profiles, and immunoglobulin heavy chain gene usage.

Aoun P, Zhou G, Chan WC, Page C, Neth K, Pickering D, Sanger W, Quinn-Laquer B, Watson P, Lynch JF, Lynch HT, Weisenburger DD
American journal of clinical pathology 2007 Jan;127(1):31-8

Modern prognostic factors and angiogenesis in chronic lymphocytic leukemia: more data needed.

Smolej L, Andrys C, Vroblova V
Leukemia research 2007 Dec;31(12):1763-4

The effect of immunoglobulin VH gene mutation status and other prognostic factors on the incidence of major infections in patients with chronic lymphocytic leukemia.

Francis S, Karanth M, Pratt G, Starczynski J, Hooper L, Fegan C, Pepper C, Valcarcel D, Milligan DW, Delgado J
Cancer 2006 Sep 1;107(5):1023-33

Feasibility of an easily applicable method of ZAP-70 measurement in chronic lymphocytic leukemia in the routine flow cytometry setting: A methodological approach.

Passam F, Tachynopoulou V, Skoumi D, Tsompanakou A, Stavropoulos-Giokas A, Vadikolia C, Anagnostopoulos A, Paterakis G
Annals of hematology 2006 Nov;85(11):795-805

A scoring system based on the expression of six surface molecules allows the identification of three prognostic risk groups in B-cell chronic lymphocytic leukemia.

Zucchetto A, Bomben R, Dal Bo M, Sonego P, Nanni P, Rupolo M, Bulian P, Dal Maso L, Del Poeta G, Del Principe MI, Degan M, Gattei V
Journal of cellular physiology 2006 May;207(2):354-63

Interleukin-21 receptor (IL-21R) is up-regulated by CD40 triggering and mediates proapoptotic signals in chronic lymphocytic leukemia B cells.

de Totero D, Meazza R, Zupo S, Cutrona G, Matis S, Colombo M, Balleari E, Pierri I, Fabbi M, Capaia M, Azzarone B, Gobbi M, Ferrarini M, Ferrini S
Blood 2006 May 1;107(9):3708-15

Comparison of bone marrow and peripheral blood ZAP-70 status examined by flow cytometric immunophenotyping in patients with chronic lymphocytic leukemia.

Sheridan R, Mounajjed T, Ehrmann DE, Hurtubise PE, Schrager JA
Cytometry. Part B, Clinical cytometry 2006 Jul 15;70(4):320-1

Evaluation of ZAP-70 expression by flow cytometry in chronic lymphocytic leukemia: A multicentric international harmonization process.

Letestu R, Rawstron A, Ghia P, Villamor N, Boeckx N, Boettcher S, Buhl AM, Duerig J, Ibbotson R, Kroeber A, Langerak A, Le Garff-Tavernier M, Mockridge I, Morilla A, Padmore R, Rassenti L, Ritgen M, Shehata M, Smolewski P, Staib P, Ticchioni M, Walker C, Ajchenbaum-Cymbalista F
Cytometry. Part B, Clinical cytometry 2006 Jul 15;70(4):309-14

Comparison of methods for determining zeta-chain associated protein - 70 (ZAP-70) expression in patients with B-cell chronic lymphocytic leukemia (B-CLL).

Bojarska-Junak A, Giannopoulos K, Kowal M, Dmoszyńska A, Roliński J
Cytometry. Part B, Clinical cytometry 2006 Jul 15;70(4):293-301

ZAP-70 expression in B-cell chronic lymphocytic leukemia: evaluation by external (isotypic) or internal (T/NK cells) controls and correlation with IgV(H) mutations.

Zucchetto A, Bomben R, Bo MD, Nanni P, Bulian P, Rossi FM, Del Principe MI, Santini S, Del Poeta G, Degan M, Gattei V
Cytometry. Part B, Clinical cytometry 2006 Jul 15;70(4):284-92

An optimized whole blood method for flow cytometric measurement of ZAP-70 protein expression in chronic lymphocytic leukemia.

Shankey TV, Forman M, Scibelli P, Cobb J, Smith CM, Mills R, Holdaway K, Bernal-Hoyos E, Van Der Heiden M, Popma J, Keeney M
Cytometry. Part B, Clinical cytometry 2006 Jul 15;70(4):259-69

ZAP-70 by flow cytometry: a comparison of different antibodies, anticoagulants, and methods of analysis.

Best OG, Ibbotson RE, Parker AE, Davis ZA, Orchard JA, Oscier DG
Cytometry. Part B, Clinical cytometry 2006 Jul 15;70(4):235-41

A robust ratio metric method for analysis of Zap-70 expression in chronic lymphocytic leukemia (CLL).

Bakke AC, Purtzer Z, Leis J, Huang J
Cytometry. Part B, Clinical cytometry 2006 Jul 15;70(4):227-34

Clinical utility of assessing ZAP-70 and CD38 in chronic lymphocytic leukemia.

Rassenti LZ, Kipps TJ
Cytometry. Part B, Clinical cytometry 2006 Jul 15;70(4):209-13

ZAP-70 in CLL: towards standardization of a biomarker for patient management: history of clinical cytometry special issue.

Marti G, Orfao A, Goolsby C
Cytometry. Part B, Clinical cytometry 2006 Jul 15;70(4):197-200

ZAP-70 is expressed by normal and malignant human B-cell subsets of different maturational stage.

Scielzo C, Camporeale A, Geuna M, Alessio M, Poggi A, Zocchi MR, Chilosi M, Caligaris-Cappio F, Ghia P
Leukemia 2006 Apr;20(4):689-95

Comparison of flow cytometric methods for the measurement of ZAP-70 expression in a routine diagnostic laboratory.

Gibbs G, Bromidge T, Howe D, Hopkins J, Johnson S
Clinical and laboratory haematology 2005 Aug;27(4):258-66

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ZAP70 in Jurkat cells. Samples were separated by SDS-PAGE, blotted with (Lane 1) or without (Lane 2) a ZAP70 monoclonal antibody (Product # MA1-21350) and developed using Goat Anti-mouse IgG-Peroxidase.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Zap-70 Monoclonal Antibody (1E7.2)(Product # MA1-21350) and a 70kDa band corresponding to Zap-70 was observed across cell lines and tissue tested. Membrane enriched extracts (30 µg lysate) of Jurkat (Lane 1), MOLT-4 (Lane 2), Raji (Lane 3), Ramos (Lane 4), EL4 (Lane 5) and MEF (Lane 6). Tissue Extracts of Mouse Thymus (Lane 7) and Mouse Liver (Lane 8) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Zap-70 was performed using 70 confluent log phase Jurkat cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Zap-70 Monoclonal Antibody (1E7.2) (Product # MA1-21350) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membrane localization. Panel e represents Raji cells with no expression of Zap-70. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.