VEGF Bioassay
A Simpler, More Consistent VEGF Bioassay
- Prequalified according to ICH guidelines
- Mechanism-of-action-based assay
- No primary cell culture required
- Supplied in convenient thaw-and-use format; also available in cell propagation model (CPM) format
Catalog Number:
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Catalog Number: GA2001
Catalog Number: GA2005
Catalog Number: GA1082
Catalog Number: J2371
安定した結果が得られるシンプルな VEGF バイオアッセイ
血管内皮増殖因子 (VEGF) は上皮細胞、がん細胞あるいはマクロファージより分泌される重要なシグナル伝達タンパク質です。血管新生促進、血管透過性の亢進、腫瘍浸潤および腫瘍生存の増強、制御性T細胞における抗腫瘍反応の阻害など多くの機能を有しています。VEGF 受容体にはいくつかのタイプ（VEGFR1, VEGFR2 および VEGFR3）が存在します。VEGFR2 (KDRとしても知られる) はVEGFに対する既知の受容体細胞応答のほとんどを調節します。VEGF には4つのアイソフォーム （VEGF-121 [拡散性] , VEGF-165 [拡散性] , VEGF-189 および VEGF-206）が存在し、体内では VEGF-165 が主要なアイソフォームとして存在します。
VEGFファミリーの全てのメンバーは、受容体チロシンキナーゼ（VEGFR1 [Flt1]) および VEGFR2 [Flk1/KDR]）への結合により細胞応答を制御します。VEGF が KDRに結合すると受容体は二量体化し、リン酸化転移を介して活性化されます。
VEGF Bioassay は VEGFによるKDR (VEGFR2) の刺激と阻害をルシフェラーゼのリードアウトとして測定する細胞ベースの発光アッセイです。本アッセイは現行法の内皮細胞増殖アッセイに存在する多くの欠点をカバーしており、VEGF反応の誘導あるいは阻害を目的とした新規な生物学的療法の発見、開発に使用することができます。
アッセイの構成品
製品に含まれる VEGF Responsive Cells は凍結保存されており溶解、播種してアッセイに直ぐに利用することができます（細胞を増やす必要もありません）。
この VEGF Responsive Cells は応答配列 (RE) で駆動する luc2Pおよび外来性 VEGF受容体を発現させるために遺伝子改変が施されています。VEGF がVEGF Responsive Cells に結合すると、受容体が細胞内にシグナルを伝え発光を生じさせます。この発光はBio-Glo™ Luciferase Assay System （カタログ番号 G7940）およびGloMax® Discover System などの標準的なルミノメーターで検出、定量することができます。
ワークフロー
従来法
標準化された試薬による最適な結果
- ”解凍 ＆ アッセイ” だけのフォーマットは従来法に比べ飛躍的に時間と労力を削減し、バラつきも低減
- キットは標準化されたフォーマットとして必要なすべての試薬を供給
- ハイスループットワークフローにも容易に適応
アッセイの原理
アッセイパフォーマンスデータ
正確度 (accuracy) と 精度 (precision) はバイオアッセイやバイオロジックス開発において非常に重要です。VEGF Bioassay は正確度と 精度を兼ね備えたアッセイです（正確度とは標的または参照値に対してどれほど近かったかという度合いであり、精度は再現性の度合いです）。本アッセイは正確度 と 精度 のためのICHガイドラインに従い開発、事前承認されており、抗体医薬の研究開発に使用するバイオアッセイの様々なアプリケーションに必要とされるパフォーマンス特性を備えています。
The VEGF Bioassay Shows Precision, Accuracy and Linearity.
| Parameter | Results | |
|---|---|---|
| Accuracy | % Expected Relative Potency | % Recovery |
| 50 | 100.1 | |
| 75 | 96.67 | |
| 150 | 102.75 | |
| 200 | 107.25 | |
| Repeatability (% CV) | 100% (Reference) | 8.73 |
| Intermediate Precision (% CV) | 9.28 | |
| Linearity (r2) | 0.9984 | |
| Linearity (y = mx + b) | y = 1.1x–7.792 | |
| A 50–200% theoretical potency series of bevacizumab (anti-VEGF antibody) was analyzed in triplicate in three independent experiments performed on three days by each of two analysts (for a total of six independent experiments). Bio-Glo™ Reagent was added and luminescence quantified using the GloMax® Discover System. Data were analyzed and relative potencies calculated after parallelism determination using JMP® software. Data were generated using thaw-and-use cells. | ||
The VEGF Bioassay response to recombinant human VEGF.
VEGF Bioassay used to measure anti-VEGF antibody activity.
キットのフォーマットと関連製品
- 1X および 5X サイズより選択可能
- Recombinant VEGF の別売品
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
KDR/NFAT-RE HEK293 Cells |
GA109A | 1 × 0.5ml |
Fetal Bovine Serum |
J121A | 1 × 4ml |
DMEM with 4.5g/L Glucose, L-Glutamine, Sodium Pyruvate |
J236A | 1 × 60ml |
Bio-Glo™ Luciferase Assay Buffer |
G719A | 1 × 10ml |
Bio-Glo™ Luciferase Assay Substrate |
G720A | 1 × 1 vial |
SDS
Search for SDS分析証明書
使用制限
Not For Medical Diagnostic Use.保存条件
Please see the Protocol for detailed storage conditions.
NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. BY USE OF THIS PRODUCT, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the recipient is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the recipient with a full refund.
This product may not be further sold or transferred by the recipient and may be used only by the recipient, and then only for (1) research use, (2) discovery, development and monitoring of biologic drugs and vaccines, (3) quality assurance testing of biologic drugs and vaccines, and (4) product release assays for biologic drugs and vaccines. No other commercial use is allowed. "Commercial use” means any and all uses of this product by recipient for monetary or other consideration, including providing a service, information or data to unaffiliated third parties, and resale of this product for any use. Recipient has no right to propagate, modify, derivatize, genetically engineer or otherwise create variations of the cells or genes stably transfected within the cells. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all luminescence assays using this product or contact Promega to obtain a license for use of this product with reagents other than Promega’s. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING AS TO MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THIS PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.
HEK293 cells were obtained under license from AdVec Inc.
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
KDR/NFAT-RE HEK293 Cells |
GA109A | 5 × 0.5ml |
Fetal Bovine Serum |
J121A | 5 × 4ml |
DMEM with 4.5g/L Glucose, L-Glutamine, Sodium Pyruvate |
J236A | 5 × 60ml |
Bio-Glo™ Luciferase Assay Buffer |
G719A | 5 × 10ml |
Bio-Glo™ Luciferase Assay Substrate |
G720A | 5 × 1 vial |
SDS
Search for SDS分析証明書
使用制限
Not For Medical Diagnostic Use.保存条件
Please see the Protocol for detailed storage conditions.
NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. BY USE OF THIS PRODUCT, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the recipient is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the recipient with a full refund.
This product may not be further sold or transferred by the recipient and may be used only by the recipient, and then only for (1) research use, (2) discovery, development and monitoring of biologic drugs and vaccines, (3) quality assurance testing of biologic drugs and vaccines, and (4) product release assays for biologic drugs and vaccines. No other commercial use is allowed. "Commercial use” means any and all uses of this product by recipient for monetary or other consideration, including providing a service, information or data to unaffiliated third parties, and resale of this product for any use. Recipient has no right to propagate, modify, derivatize, genetically engineer or otherwise create variations of the cells or genes stably transfected within the cells. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all luminescence assays using this product or contact Promega to obtain a license for use of this product with reagents other than Promega’s. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING AS TO MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THIS PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.
HEK293 cells were obtained under license from AdVec Inc.
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
KDR/NFAT-RE HEK293 Cells (CPM) |
GA108A | 2 × 1ml |
SDS
Search for SDS分析証明書
使用制限
Not For Medical Diagnostic Use.保存条件
Please see the Protocol for detailed storage conditions.
NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. BY USE OF THIS PRODUCT, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the recipient is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the recipient with a full refund.
This product may not be further sold or transferred by the recipient and may be used only by the recipient, and then only for (1) research use, (2) discovery, development and monitoring of biologic drugs and vaccines, (3) quality assurance testing of biologic drugs and vaccines, and (4) product release assays for biologic drugs and vaccines. No other commercial use is allowed. "Commercial use" means any and all uses of this product by recipient for monetary or other consideration, including providing a service, information or data to unaffiliated third parties, and resale of this product for any use. Recipient has no right to modify, derivatize, genetically engineer or otherwise create variations of the cells or genes stably transfected within the cells except that recipient may propagate and store the cells for long-term use. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all luminescence assays using this product or contact Promega to obtain a license for use of this product with reagents other than Promega's. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING AS TO MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THIS PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.
HEK293 cells were obtained under license from AdVec Inc.
選択製品の構成品内容
| Item | Part # | Size | Concentration |
|---|---|---|---|
Recombinant VEGF |
J237A | 1 × 10μg | 200μg/ml |
SDS
Search for SDS分析証明書
使用制限
Not For Medical Diagnostic Use.保存条件
Please see the Protocol for detailed storage conditions.
Resources
No related resources available
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関連製品
Bio-Glo™ Luciferase Assay System
レポーターバイオアッセイに理想的な安定性の高いルシフェラーゼ試薬
G7940, G7941
GloMax® Discover System
発光・蛍光・吸光度測定に対応した高性能マイクロプレートリーダー
GM3000
RealTime-Glo™ MT Cell Viability Assay
72 時間まで培養細胞の生存性をモニタリングできる発光アッセイ
G9711, G9712, G9713
CellTiter-Glo® 2.0 Cell Viability Assay
試薬の安定性が向上したCellTiter-Glo®、ATP検出に基づく細胞増殖の定量が可能
G9241, G9242, G9243
