Pengertian ELISA
Enzyme linked immunosorbent assay biasa disingkat ELISA adalah metode yang banyak digunakan untuk mendeteksi dan mengukur protein spesifik dalam sampel kompleks. Awalnya digunakan oleh Engvall dan Perlmann (1971), metode ini memungkinkan analisis sampel protein yang diimobilisasi dalam sumur pelat mikro menggunakan antibodi spesifik.
ELISA biasanya dilakukan pada pelat polistiren 96-sumur atau 384-sumur, yang secara pasif mengikat antibodi dan protein. Pengikatan dan imobilisasi reagen inilah yang membuat ELISA mudah dirancang dan dilakukan. Reagen ELISA yang diimobilisasi ke permukaan pelat mikro memudahkan untuk memisahkan bahan yang terikat dari bahan yang tidak terikat selama pengujian. Kemampuan untuk menggunakan antibodi afinitas tinggi dan membilas bahan terikat non-spesifik menjadikan ELISA sebagai metode andal untuk mengukur analit spesifik dalam preparat kasar.
Meskipun banyak varian ELISA telah dikembangkan dan digunakan dalam kondisi yang berbeda, semuanya bergantung pada 4 elemen dasar yang sama:
● Pelapisan/penangkapan (Coating/Capture) —imobilisasi antigen secara langsung atau tidak langsung ke permukaan sumur pelat mikro polistiren.
● Blocking Pelat —penambahan protein atau molekul lain yang tidak relevan untuk menutupi semua situs pengikatan permukaan tak jenuh dari sumur pelat mikro.
● Probing/deteksi—inkubasi dengan antibodi-spesifik antigen yang berikatan dengan antigen.
● Pengukuran sinyal—deteksi sinyal yang dihasilkan langsung (direct) atau sekunder pada antibodi spesifik.
Berikut merupakan perbedaan format ELISA kit komersial yang tersedia di pasaran. Ada yang hanya berupa antibody pairs, yaitu pasangan antibodi primer dan sekunder yang telah dioptimasi untuk ELISA. Juga ada ELISA kit yang pengerjaannya di bawah 3 jam saja, misalnya saja Quikey ELISA kit dari Elabscience.
Tabel perbedaan format ELISA kit
Antibody pair kits | Uncoated ELISA kits | Coated ELISA kits | Quickey ELISA kits | |
Butuh pelapisan pelat | Ya, inkubasi semalam | Ya, inkubasi semalam | Tidak | Tidak |
Waktu Inkubasi | 24 jam | 24 jam | 2.5–4 jam | 3 jam
/td> |
Pengerjaan | 1 jam 30 menit | 1 jam 30 menit | 1 jam | 40 menit |
Readout
|
HRP-TMB (colorimetric) | HRP-TMB (colorimetric) | HRP-TMB (colorimetric) | HRP-TMB (colorimetric) |
Alat Baca
|
Microplate reader, absorbans | Microplate reader, absorbans | Microplate reader, absorbans | Microplate reader, absorbans |
Waktu pembacaan | 2 min | 2 min | 2 min | 2 min |
ELISA dirancang untuk mengukur jumlah target yang terikat antara pasangan antibodi yang cocok (matched antibody pair). Antibodi spesifik target dilapisi ke dasar sumur pelat mikro, biasanya membutuhkan waktu inkubasi semalam. Sampel, standar, atau kontrol kemudian ditambahkan ke dalam sumur-sumur ini dan mengikat antibodi yang diimobilisasi (capture antibody). Sandwich dibentuk dengan penambahan antibodi kedua (detection antibody), larutan substrat ditambahkan yang bereaksi dengan kompleks enzim-antibodi-target untuk menghasilkan sinyal yang terukur. Intensitas sinyal ini berbanding lurus dengan konsentrasi target yang ada dalam spesimen asli.
Perbedaan Coated dan Uncoated ELISA Kit
Coated ELISA kit
Coated ELISA kit adalah kit ELISA siap pakai (ready to use) yang tersedia secara komersial untuk mendeteksi ratusan sitokin, kemokin, faktor pertumbuhan, analit neurobiologi dan target lainnya dalam berbagai area penelitian. Coated ELISA kit ini berisi semua reagen lengkap yang dibutuhkan untuk menjalankan prosedur ELISA. Reagen ELISA tersebut di antaranya adalah pelat antibodi pra-pelapisan, antibodi pendeteksi, buffer, pengencer, standar dan substrat.
Berikut contoh komponen ELISA kit Elabscience Human Interleukin 8 (IL-8) nomor katalog E-EL-H6008.
Uncoated ELISA kit
Uncoated ELISA merupakan format ELISA yang dirancang untuk para peneliti yang dananya terbatas namun sampelnya banyak. Memang prosesnya memakan waktu lebih lama dibanding pengerjaan ELISA ready to use, namun itu semua sebanding dengan harga yang lebih rendah dan juga dimungkinkan modifikasi uji ELISA yang dilakukan.
Sebagai contoh Elabscience Human Interleukin 8 (IL-8) Uncoated ELISA Kit catalog number: E-UNEL-H0099, berisi pasangan antibodi yang telah dicocokkan sebelumnya, dan reagen untuk melakukan uji ELISA untuk mendeteksi dan mengukur kadar protein IL-8 pada manusia.
Jika order uncoated ELISA kit, pelanggan direkomendasikan untuk sekalian order Ancillary Reagent Kit (Cat No. E-ELIR-K001), yang berisi semua reagen yang dibutuhkan selain pada gambar 2 di atas. Ancillary Reagent Kit ini cukup untuk pengerjaan 5×96 test ELISA.
Marker Uncoated ELISA Kit Best Seller
Selain Elabscience terdapat pula beberapa merk lainnya sebagai manufaktur uncoated ELISA kit misalnya saja Invitrogen (grup Thermo). Hampir semua ELISA kit yang dijual oleh merk Invitrogen berikut, ada juga pada merk Elabscience.
● iga mouse uncoated elisa kit (Elabscience; E-UNEL-H0070)
● invitrogen mouse il-6 uncoated elisa (Elabscience; E-UNEL-M0070)
● invitrogen mouse tnf alpha uncoated elisa kit (Elabscience; E-UNEL-M0103)
● human ifn gamma uncoated elisa invitrogen (Elabscience; E-UNEL-M0043)
● il-2 human uncoated elisa kit (Elabscience; E-UNEL-M0065)
● invitrogen human tnf alpha uncoated elisa (Elabscience; E-UNEL-H0175)
● il-8 human uncoated elisa kit (Elabscience; E-UNEL-H0099)
● igg (total) human uncoated elisa kit (Elabscience; E-UNEL-H0079)
Untuk spesifikasi, kedua merk hampir sama. Namun, dengan kualitas hampir setara, harga Elabscience ELISA kit lebih terjangkau, pengiriman lebih cepat dan kisaran produk lebih lengkap. Selain itu juga, produk Elabscience sudah banyak digunakan oleh para peneliti pada jurnal-jurnal ilmiah kelas dunia.
Berikut merupakan list uncoated ELISA kit merk Elabscience.
Human Uncoated ELISA Kit (5x96T; 15x96T)
Catalog Number | Target Species | Product Name |
E-UNEL-H0003 | Human | Uncoated Human ADP/Acrp30(Adiponectin) ELISA Kit |
E-UNEL-H0006 | Human | Uncoated Human ALB(Albumin) ELISA Kit |
E-UNEL-H0012 | Human | Uncoated Human ApoB(Apolipoprotein B) ELISA Kit |
E-UNEL-H0015 | Human | Uncoated Human BDNF(Brain Derived Neurotrophic Factor) ELISA Kit |
E-UNEL-H0021 | Human | Uncoated Human C3a(Complement Component 3a) ELISA Kit |
E-UNEL-H0022 | Human | Uncoated Human C5a(Complement Component 5a) ELISA Kit |
E-UNEL-H0025 | Human | Uncoated Human CEA(Carcinoembryonic Antigen) ELISA Kit |
E-UNEL-H0027 | Human | Uncoated Human CHEM(Chemerin) ELISA Kit |
E-UNEL-H0029 | Human | Uncoated Human CRP(C-Reactive Protein) ELISA Kit |
E-UNEL-H0031 | Human | Uncoated Human CTSB(Cathepsin B) ELISA Kit |
E-UNEL-H0035 | Human | Uncoated Human Cys-C(Cystatin C) ELISA Kit |
E-UNEL-H0036 | Human | Uncoated Human D2D(D-Dimer) ELISA Kit |
E-UNEL-H0038 | Human | Uncoated Human DPP4(Dipeptidyl Peptidase Ⅳ) ELISA Kit |
E-UNEL-H0040 | Human | Uncoated Human EGF(Epidermal Growth Factor) ELISA Kit |
E-UNEL-H0041 | Human | Uncoated Human EGFR(Epidermal Growth Factor Receptor) ELISA Kit |
E-UNEL-H0042 | Human | Uncoated Human EMMPRIN/CD147(Extracellular Matrix Metalloproteinase Inducer) ELISA Kit |
E-UNEL-H0044 | Human | Uncoated Human EPO(Erythropoietin) ELISA Kit |
E-UNEL-H0050 | Human | Uncoated Human FE(Ferritin) ELISA Kit |
E-UNEL-H0057 | Human | Uncoated Human GDF15(Growth Differentiation Factor 15) ELISA Kit |
E-UNEL-H0060 | Human | Uncoated Human GH(Growth Hormone) ELISA Kit |
E-UNEL-H0063 | Human | Uncoated Human Hepc(Hepcidin) ELISA Kit |
E-UNEL-H0064 | Human | Uncoated Human HGF(Hepatocyte Growth Factor) ELISA Kit |
E-UNEL-H0066 | Human | Uncoated Human ICAM-1/CD54(intercellular adhesion molecule 1) ELISA Kit |
E-UNEL-H0067 | Human | Uncoated Human IFABP/FABP2(Intestinal Fatty Acid Binding Protein) ELISA Kit |
E-UNEL-H0068 | Human | Uncoated Human IFN-β(Interferon Beta) ELISA Kit |
E-UNEL-H0069 | Human | Uncoated Human IFN-γ(Interferon Gamma) ELISA Kit |
E-UNEL-H0070 | Human | Uncoated Human IgA(Immunoglobulin A) ELISA Kit |
E-UNEL-H0072 | Human | Uncoated Human IGF-1(Insulin-like Growth Factor 1) ELISA Kit |
E-UNEL-H0079 | Human | Uncoated Human IgG(Immunoglobulin G) ELISA Kit |
E-UNEL-H0084 | Human | Uncoated Human IL-10(Interleukin 10) ELISA Kit |
E-UNEL-H0085 | Human | Uncoated Human IL-12/P70(Interleukin 12/P70) ELISA Kit |
E-UNEL-H0088 | Human | Uncoated Human IL-18(Interleukin 18) ELISA Kit |
E-UNEL-H0092 | Human | Uncoated Human IL-2(Interleukin 2) ELISA Kit |
E-UNEL-H0096 | Human | Uncoated Human IL-4(Interleukin 4) ELISA Kit |
E-UNEL-H0097 | Human | Uncoated Human IL-6(Interleukin 6) ELISA Kit |
E-UNEL-H0099 | Human | Uncoated Human IL-8(Interleukin 8) ELISA Kit |
E-UNEL-H0100 | Human | Uncoated Human INHB(Inhibin B) ELISA Kit |
E-UNEL-H0101 | Human | Uncoated Human INS(Insulin) ELISA Kit |
E-UNEL-H0104 | Human | Uncoated Human LBP(Lipopolysaccharide Binding Protein) ELISA Kit |
E-UNEL-H0105 | Human | Uncoated Human LEP(Leptin) ELISA Kit |
E-UNEL-H0109 | Human | Uncoated Human LpPLA2(Lipoprotein-associated Phospholipase A2) ELISA Kit |
E-UNEL-H0112 | Human | Uncoated Human MCP-1(Monocyte Chemotactic Protein 1) ELISA Kit |
E-UNEL-H0115 | Human | Uncoated Human MMP-1(Matrix Metalloproteinase 1) ELISA Kit |
E-UNEL-H0116 | Human | Uncoated Human MMP-3(Matrix Metalloproteinase 3) ELISA Kit |
E-UNEL-H0117 | Human | Uncoated Human MMP-8(Matrix Metalloproteinase 8) ELISA Kit |
E-UNEL-H0118 | Human | Uncoated Human MMP-9(Matrix Metalloproteinase 9) ELISA Kit |
E-UNEL-H0120 | Human | Uncoated Human MUC1(Mucin 1) ELISA Kit |
E-UNEL-H0123 | Human | Uncoated Human NGAL(Neutrophil Gelatinase Associated Lipocalin) ELISA Kit |
E-UNEL-H0127 | Human | Uncoated Human NSE(Neuron Specific Enolase) ELISA Kit |
E-UNEL-H0130 | Human | Uncoated Human OPG(Osteoprotegerin) ELISA Kit |
E-UNEL-H0134 | Human | Uncoated Human PCSK9(Proprotein Convertase Subtilisin/Kexin Type 9) ELISA Kit |
E-UNEL-H0149 | Human | Uncoated Human RETN(Resistin) ELISA Kit |
E-UNEL-H0150 | Human | Uncoated Human sCD14(Soluble Cluster of Differentiation 14) ELISA Kit |
E-UNEL-H0155 | Human | Uncoated Human SDF-1/CXCL12(Stromal Cell Derived Factor 1) ELISA Kit |
E-UNEL-H0156 | Human | Uncoated Human SELE(E-Selectin) ELISA Kit |
E-UNEL-H0165 | Human | Uncoated Human TF(Transferrin) ELISA Kit |
E-UNEL-H0169 | Human | Uncoated Human TGF-β1(Transforming Growth Factor Beta 1) ELISA Kit |
E-UNEL-H0171 | Human | Uncoated Human TIMP-1(Tissue Inhibitors of Metalloproteinase 1) ELISA Kit |
E-UNEL-H0175 | Human | Uncoated Human TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit |
E-UNEL-H0179 | Human | Uncoated Human VEGF165(Vascular Endothelial Growth Factor 165) ELISA Kit |
E-UNEL-H0186 | Human | Uncoated Human αFP(Alpha-Fetoprotein) ELISA Kit |
Mouse Uncoated ELISA Kit (5x96T; 15x96T)
Catalog Number | Target Species | Product Name |
E-UNEL-M0004 | Mouse | Uncoated Mouse ADP/Acrp30(Adiponectin) ELISA Kit |
E-UNEL-M0016 | Mouse | Uncoated Mouse CRP(C-Reactive Protein) ELISA Kit |
E-UNEL-M0021 | Mouse | Uncoated Mouse Cys-C(Cystatin C) ELISA Kit |
E-UNEL-M0023 | Mouse | Uncoated Mouse EGF(Epidermal Growth Factor) ELISA Kit |
E-UNEL-M0043 | Mouse | Uncoated Mouse IFN-γ(Interferon Gamma) ELISA Kit |
E-UNEL-M0044 | Mouse | Uncoated Mouse IgA(Immunoglobulin A) ELISA Kit |
E-UNEL-M0045 | Mouse | Uncoated Mouse IGF-1(Insulin-like Growth Factor 1) ELISA Kit |
E-UNEL-M0050 | Mouse | Uncoated Mouse IgG(Immunoglobulin G) ELISA Kit |
E-UNEL-M0058 | Mouse | Uncoated Mouse IL-12 p40(Interleukin 12 p40) ELISA Kit |
E-UNEL-M0059 | Mouse | Uncoated Mouse IL-12(Interleukin 12) ELISA Kit |
E-UNEL-M0061 | Mouse | Uncoated Mouse IL-17A(Interleukin 17A) ELISA Kit |
E-UNEL-M0064 | Mouse | Uncoated Mouse IL-1β(Interleukin 1 Beta) ELISA Kit |
E-UNEL-M0065 | Mouse | Uncoated Mouse IL-2(Interleukin 2) ELISA Kit |
E-UNEL-M0068 | Mouse | Uncoated Mouse IL-4(Interleukin 4) ELISA Kit |
E-UNEL-M0070 | Mouse | Uncoated Mouse IL-6(Interleukin 6) ELISA Kit |
E-UNEL-M0072 | Mouse | Uncoated Mouse KIM-1(Kidney Injury Molecule 1) ELISA Kit |
E-UNEL-M0074 | Mouse | Uncoated Mouse LEP(Leptin) ELISA Kit |
E-UNEL-M0076 | Mouse | Uncoated Mouse LTF/LF(Lactoferrin) ELISA Kit |
E-UNEL-M0077 | Mouse | Uncoated Mouse MCP-1(Monocyte Chemotactic Protein 1) ELISA Kit |
E-UNEL-M0085 | Mouse | Uncoated Mouse OPG(Osteoprotegerin) ELISA Kit |
E-UNEL-M0089 | Mouse | Uncoated Mouse Pro-MMP-9(Pro-Matrix Metalloproteinase-9) ELISA Kit |
E-UNEL-M0099 | Mouse | Uncoated Mouse TGF-β1(Transforming Growth Factor Beta 1) ELISA Kit |
E-UNEL-M0103 | Mouse | Uncoated Mouse TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit |
E-UNEL-M0106 | Mouse | Uncoated Mouse VEGF-A(Vascular Endothelial Cell Growth Factor A) ELISA Kit |
Rat Uncoated ELISA Kit (5x96T; 15x96T)
Catalog Number | Target Species | Product Name |
E-UNEL-R0002 | Rat | Uncoated Rat ADP/Acrp30(Adiponectin) ELISA Kit |
E-UNEL-R0004 | Rat | Uncoated Rat BMG/β2-MG(Beta-2-Microglobulin) ELISA Kit |
E-UNEL-R0006 | Rat | Uncoated Rat CRP(C-Reactive Protein) ELISA Kit |
E-UNEL-R0019 | Rat | Uncoated Rat ICAM-1/CD54(Intercellular Adhesion Molecule 1) ELISA Kit |
E-UNEL-R0021 | Rat | Uncoated Rat IgG(Immunoglobulin G) ELISA Kit |
E-UNEL-R0023 | Rat | Uncoated Rat IL-10(Interleukin 10) ELISA Kit |
E-UNEL-R0024 | Rat | Uncoated Rat IL-18(Interleukin 18) ELISA Kit |
E-UNEL-R0028 | Rat | Uncoated Rat IL-1β(Interleukin 1 Beta) ELISA Kit |
E-UNEL-R0029 | Rat | Uncoated Rat IL-2(Interleukin 2) ELISA Kit |
E-UNEL-R0030 | Rat | Uncoated Rat IL-6(Interleukin 6) ELISA Kit |
E-UNEL-R0033 | Rat | Uncoated Rat KIM-1(Kidney Injury Molecule 1) ELISA Kit |
E-UNEL-R0039 | Rat | Uncoated Rat MMP-9(Matrix Metalloproteinase 9) ELISA Kit |
E-UNEL-R0044 | Rat | Uncoated Rat PCSK9(Proprotein convertase subtilisin/kexin type 9) ELISA Kit |
E-UNEL-R0046 | Rat | Uncoated Rat PRL(Prolactin) ELISA Kit |
E-UNEL-R0054 | Rat | Uncoated Rat TGF-β1(Transforming Growth Factor Beta 1) ELISA Kit |
E-UNEL-R0055 | Rat | Uncoated Rat TIMP-1(Tissue Inhibitors of Metalloproteinase 1) ELISA Kit |
E-UNEL-R0057 | Rat | Uncoated Rat TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit |