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9+ +10.+Western+Blot+Solution KR Compressed Compressed2 Compressed+PDF - Io
9+ +10.+Western+Blot+Solution KR Compressed Compressed2 Compressed+PDF - Io
9+ +10.+Western+Blot+Solution KR Compressed Compressed2 Compressed+PDF - Io
위한 최상의 솔루션
Customize your western blot tools to
optimize your outcomes
웨스턴 블랏 워크플로우의 모든 단계에서 당신에게 적합한 솔루션을 찾아보세요.
그 솔루션은 뛰어난 결과를 가져다주며 당신의 시간과 노력을 아낄 수 있습니다.
–
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Separate Transfer
Page
Precast protein gels 8, 37 Fast protein transfer systems
Mini gel system 9 Transfer membranes
Power supplies 18 Wet transfer
Midi gel system 20
Large-volume gel running buffers 21
Fluorescence-compatible sample buffers 26
Protein ladders 21, 27
Handcast protein gel system 36
2
+
Detect
Page Page
10, 22 Automated western blot processing systems 12, 23
28 Western blot and gel imaging systems 14, 32
40 Blocking buffers 29, 41
Fluorescent secondary antibodies 30
Chemiluminescent substrates 42
Stripping buffers 45
3
Contents
High-efficiency western blotting Multiplex western blotting
Separate Separate
Precast gels 8 Fluorescence-compatible
Mini gel tank 9 sample buffers 26
Prestained fluorescent
Transfer protein ladders 27
Fast protein transfer 10
Transfer
Detect Fluorescence-friendly
Automated western blot transfer membranes 28
processing systems 12
Detect
Western blot and gel imaging 14
High-quality, filtered blocking buffers 29
Ordering information 16
Fluorescent secondary antibodies 30
Fluorescent imaging 32
High-throughput western blotting
Ordering information 33
Separate
Power supplies 18
Optimizing your western blots
Midi gel systems 20
Separate
Large-volume buffers 21
Handcast system 36
Transfer Protein gels and buffers 37
Flexible protein transfer 22
Transfer
Detect Wet transfer 40
Automated western blot processing 23
Detect
Ordering information 24
Blocking buffers 41
Chemiluminescent detection 42
Stripping buffers 45
Ordering information 46
High-efficiency western blotting
High-efficiency
western blotting
당신의 시간은 우리에게 소중합니다.
우리는 당신이 시간을 아껴 웨스턴 블랏의
결과를 보다 빠르게 볼 수 있도록 지원하고
있습니다.
Invitrogen™ iWestern™ workflow는
혁신적이고 모던한 솔루션이며
당신의 시간과 노력을 아낄 수 있게
고안되었습니다. Precast gel부터 자동
detection 시스템까지 우리는 고품질의
웨스턴 블랏 제품들로 빠르게 결과를 볼 수
있는 솔루션을 제공합니다.
5
Separate Transfer Detect
Invitrogen
Invitrogen ™
Bolt™ Bis-Tris
Bolt Bis-Tris Plus
Plus Gels
Gels Invitrogen
Invitrogen ™
iBlot™2 2Dry
GelTransfer
TransferDevice
Device Invitrogen WesternSystems
Invitrogen™ iBind ™Western Systems Invitrogen
Invitrogen
™
ImagingSystems
iBright™Imaging Systems
with Invitrogen™ Mini Gel Tank
Compare workflow and data obtained using the iWestern workflow bundle and manual
western blotting. The iWestern workflow enables the detection of HDAC1 in HeLa lysate in
approximately 4 hr vs. 18 hr using the manual western workflow. Two-fold serial dilutions (starting
with 20 µg of HeLa lysate) were loaded and separated on Tris-glycine gels. The proteins were
transferred to nitrocellulose membranes and detected using Thermo Scientific™ SuperSignal™ West
Pico PLUS chemiluminescent substrate.
6
Separate Transfer Detect
• Invitrogen™ Protein Gel Welcome Pack • Invitrogen™ iBind™ Western Starter Kit
(including Mini Gel Tank)
• Invitrogen™ iBright™ FL1500 Imaging System
• Invitrogen™ iBlot™ 2 Starter Kit
7
Separate Transfer Detect
MagicMark standard
precast polyacrylamide gel로 denaturing 조건에서
폭 넓은 범위의 크기를 가진 단백질 분리에 최적으로
MAPK14
MAPK14
EPHB3
EPHB3
DDR2
DDR2
FLT1
FLT1
IKKβ
IKKβ
HCK
HCK
GST
GST
• 샘플 로딩양 확대 – 최대 60 μL 수준의 샘플을 다룰 수 Bolt Bis-Tris Plus gel Bio-Rad TGX gel
있도록 만들어진 WedgeWell은 희석된 샘플이나 적게 Bolt Bis-Tris Plus mini gels help provide better western
발현된 단백질 샘플도 쉽게 검출할 수 있게 합니다. blotting results. A western blot of a Bolt gel shows clean,
sharp protein signals corresponding to only full-length proteins,
whereas a western blot of a Bio-Rad™ TGX™ gel shows multiple
• Lot-to-lot의 높은 일관성 – Rf 값의 변동계수(CV)가
low molecular weight degradation products.
단 2% 수준 (migration)
• 개선된 밴드의 품질 - 선명한 직선 밴드를 확인할 수 • Mini Gel Tank는 기존의 전기 영동 탱크보다 직관적이며
있습니다. 사용하기 편리한 구조로 만들어졌습니다.
8
Separate Transfer Detect
9
Separate Transfer Detect
Invitrogen™ iBlot™ 2 Gel Transfer Device는 iBlot 2 Starter Kit (Cat. No. IB21001S)
프리미엄 웨스턴 블랏 트랜스퍼 장치로 높은 퍼포먼스와
편의성을 가지고 있습니다. iBlot2 장치는 조립된 트랜스퍼
스택을 이용해 최적화된 혁신적인 시스템을 이용하는 • 간소화된 준비와 정리 - iBlot2 스택은 트랜스퍼 세팅과
유일한 장치입니다. 어지럽게 트랜스퍼 버퍼를 준비할 정리 작업을 간편하게 합니다.
필요없이 스택에 Gel만 넣고 장치를 작동시키세요.
• 편의성 - 터치 스크린 인터페이스와 최적화된 트랜스퍼
• 빠른 트랜스퍼 - 전극 간의 거리를 좁히고 높은 전류와 프로토콜, 패킹된 스택이 Midi gel 또는 Mini gel을
자기장을 형성해 트랜스퍼 시간을 7분으로 단축시킵니다. 위해 준비되어 있습니다.
thermofisher.com/iblot2에서
더 자세한 내용을 확인해보세요
A B C
D E F
Bolt Bis-Tris Plus, PVDF NuPAGE Bis-Tris, PVDF Novex Tris-glycine, PVDF
Membranes processed on the iBlot 2 Gel Transfer Device show consistent transfer across various protein gel chemistries
to both nitrocellulose (NC) and PVDF membranes. Total cell extracts from A431 cells were transferred to NC (A–C) or PVDF
(D–F) membranes from Invitrogen™ Bolt™ 4–12% Bis-Tris gels, Invitrogen™ NuPAGE™ 4–20% Bis-Tris gels, and Invitrogen™ Novex™
4–20% Tris-glycine gels.
10
Separate Transfer Detect
EGFR
Hsp90
Tubulin
Traditional wet tank iBlot 2 system
60 min transfer 7 min transfer
p23
A431 lysate
iBlot 2 system
EGFR
7 min transfer
Hsp90
PDI EGFR
Hsp90
Tubulin
p23
p23
Chemiluminescent detection: high transfer efficiencies Multiplex fluorescent detection: high transfer efficiencies
achieved using the iBlot 2 Dry Blotting System. A431 achieved using the iBlot 2 Dry Blotting System. A431 lysate
lysate was separated on Invitrogen™ Novex™ Tris-glycine gels was separated on NuPAGE Bis-Tris gels and transferred to
and transferred to nitrocellulose using a traditional wet tank nitrocellulose using a traditional wet tank system or the iBlot 2
system or the iBlot 2 Gel Transfer Device. The membrane Gel Transfer Device. The membrane was probed for EGFR,
was probed for EGFR, Hsp90, PDI, and p23 and detected Hsp90, tubulin, and p23.
using Thermo Scientific™ SuperSignal™ West Dura Extended
Duration Substrate.
11
Separate Transfer Detect
12
Separate Transfer Detect
Fluorescence detection with the iBind Flex Western Device. Comparison of strip blots processed manually (probing and
washing steps performed in a tray) vs. with the iBind Flex Western Device. Blots were produced by separating samples on Bolt
4–12%, 10-well gels with MES SDS running buffer, rapid-dry transfer to nitrocellulose membrane using the iBlot 2 system and then
cutting each into 3-lane strips. Get more information about the target proteins and antibodies used at thermofisher.com/ibind.
13
Separate Transfer Detect
Simplified western blot and gel imaging iBright Imaging Systems provide:
Invitrogen™ iBright™ Imaging Systems으로 보다 • 버튼만 누르면, 노출 시간 최적화 - Smart Exposure
쉽고 빠르게 젤과 웨스턴 블랏을 캡쳐하고 분석해보세요. 는 반복적인 노출을 최소화하기 위해 신호에서
직관적인 인터페이스와 워크플로우가 당신의 경험치를 요구하는 최적의 노출 시간을 빠르게 결정할 수 있도록
올려줍니다. 도와줍니다.
14
Separate Transfer Detect
iBright 이미징 시스템은 당신의 다양한 어플리케이션을 지원하기 위한 5가지 이미징 모드를
제공합니다. Protein gel, nucleic acid gel, chemiluminescent 블랏, 형광 블랏 그리고
그 이상의 이미지를 간편하고 효율적으로 캡쳐할 수 있도록 합니다.
Fluorescent western blots Chemiluminescent Combined fluorescent Colorimetric western Colorimetric stained
western blots and chemiluminescent blots protein gels
western blots
Images pictured for fluorescent stained nucleic acid gels and colorimetric stained protein gels shown in pseudocolor
(false color applied). Data is captured in grayscale.
15
Ordering information
Separate Quantity Cat. No.
Mini Gel Tank 1 unit A25977
Mini Gel Tank and Blot Module Set 1 set NW2000
Bolt Bis-Tris 4–12%, 10-well Welcome Pack 1 kit NW0412A
Bolt Bis-Tris 4–12%, 15-well Welcome Pack 1 kit NW0412B
Transfer Quantity Cat. No.
iBlot 2 Gel Transfer Device 1 device IB21001
iBlot 2 Starter Kit 1 kit IB21001S
iBlot 2 Transfer Stacks, Nitrocellulose, Regular 10 stacks IB23001
iBlot 2 Transfer Stacks, Nitrocellulose, Mini 10 stacks IB23002
iBlot 2 Transfer Stacks, PVDF, Regular 10 stacks IB24001
iBlot 2 Transfer Stacks, PVDF, Mini 10 stacks IB24002
Bolt Welcome Pack with iBlot 2 Dry Blotting System 1 kit NW0412AIB2
Detect Quantity Cat. No.
iBind Western Starter Kit 1 kit SLF1000S
iBind Western Device 1 device SLF1000
10 cards SLF1010
iBind Cards
40 cards SLF1010X4
1 kit SLF1020
iBind Solution Kit
4 kits SLF1020X4
iBind Flex Western Starter Kit 1 kit SLF2000S
1 device SLF2000
iBind Flex Western Device
2 devices SLF20002PK
10 cards SLF2010
iBind Flex Cards
40 cards SLF2010X4
1 kit SLF2020
iBind Flex Solution Kit
4 kits SLF2020X4
iBright CL750 Imaging System 1 instrument A44116
iBright CL1500 Imaging System 1 instrument A44114
iBright FL1500 Imaging System 1 instrument A44115
16
High-throughput western blotting
High-throughput
western blotting
Processing many samples or blots?
우리는 당신의 대량 처리(High-throughput)
실험에 필요한 제품들을 제공하고 있습니다.
최대 26개의 샘플을 로딩할 수 있는 Precast
midi gel부터 가장 일반적으로 사용하는 대용량
버퍼까지, 우리는 당신이 필요한 웨스턴 블랏의
모든 것을 가지고 있습니다.
17
Separate Transfer Detect
• 정전압, 전류 또는 전원 설정
• 정전압, 전류 또는 전원 설정
18
Separate Transfer Detect
Load up to 26 samples
19
Separate Transfer Detect
20
Separate Transfer Detect
Bulk reagents
—~235—
—~120— —~250—
—~180—
550 nm and 700 nm fluorescent detection
—~130—
—~80— —~130—
—~70— —~100— —~100—
—~70— —~70—
—~50—
—~15—
Visible light
Visible light
—~10— —~10—
—~10—
21
Separate Transfer Detect
The Power Blotter offers high transfer efficiency for a broad range of protein sizes
Power Blotter with Pre-cut Membranes and Filters
Apparatus and consumables Power Blotter with Select Transfer Stacks
and Power Blotter 1-Step Transfer Buffer
Transfer time 5–10 min 5–10 min
KLH KLH
EGFR EGFR
Hsp90 Hsp90
PDI PDI
EGFR EGFR
Transfer efficiency (extra band) (extra band)
Cyclophilin B Cyclophilin B
Invitrogen™ Power Blotter Select Transfer Stacks and Power Blotter Pre-cut Membranes and Filters efficiently transfer high,
medium, and low molecular weight proteins. Western blot analysis of several targets (KLH, EGFR, Hsp90, PDI, and cyclophilin B
protein) was performed by loading serially diluted HeLa cell lysate with KLH spike (starting at 7.5 µg HeLa lysate, 7.5 µg KLH spike per well,
serially diluted 2:3) onto Bolt 4–12% Bis-Tris Plus gels. Proteins were transferred in 7 min using a Power Blotter Select Transfer Stack (left,
Cat. No. PB5310 and PB3310) or Power Blotter Pre-cut Membranes and Filters (right, Cat. No. PB9320 and PB7320), probed with target-
specific primary antibodies and fluorescently conjugated secondary antibodies. Images were captured using automatic exposure on an
iBright FL1000 Imaging System.
22
Separate Transfer Detect
Comparison of mini blots processed with the Bandmate Automated Western Blot Processor vs. manual processing (probing
and washing steps performed in a tray and on a shaker platform). The blots processed with the Bandmate device show comparable
intensity levels across multiple probed targets compared to the blots processed with a manual processing procedure.
23
Ordering information
Separate Quantity Cat. No.
PowerEase Touch 350W Power Supply (230 VAC) 1 unit PS0351
PowerEase Touch 120W Power Supply (230 VAC) 1 unit PS0121
XCell4 SureLock Midi-Cell 1 unit WR0100
SureLock Tandem Midi Gel Tank 1 unit STM1001
Novex Tris-Glycine SDS Running Buffer 5L LC26755
NuPAGE MES SDS Running Buffer 5L NP000202
NuPAGE MOPS SDS Running Buffer 5L NP000102
Spectra Multicolor Broad Range Protein Ladder 10 x 250 µL 26623
PageRuler Plus Prestained Protein Ladder 10 x 250 µL 26620
PageRuler Prestained Protein Ladder, 10 to 180 kDa 10 x 250 µL 26617
SimplyBlue SafeStain 3.5 L LC6065
GelCode Blue Stain Reagent 3.5 L 24592
Transfer Quantity Cat. No.
Power Blotter Welcome Pack 1 kit PB0112
Power Blotter XL Welcome Pack 1 kit PB0113
Power Blotter Station 1 unit PB0010
Power Blotter Cassette 1 cassette PB0002
Power Blotter Cassette XL 1 cassette PB0003
Power Blotter System 1 system PB0012
Power Blotter XL System 1 system PB0013
10 stacks PB3210
Power Blotter Select Transfer Stacks, nitrocellulose, mini
40 stacks PB3240
10 stacks PB5210
Power Blotter Select Transfer Stacks, PVDF, mini
40 stacks PB5240
10 stacks PB3310
Power Blotter Select Transfer Stacks, nitrocellulose, regular size
40 stacks PB3340
10 stacks PB5310
Power Blotter Select Transfer Stacks, PVDF, regular size
40 stacks PB5340
Power Blotter Pre-cut Membranes and Filters, nitrocellulose, mini 20 stacks PB7220
Power Blotter Pre-cut Membranes and Filters, PVDF, mini 20 stacks PB9220
Power Blotter Pre-cut Membranes and Filters, nitrocellulose, regular size 20 stacks PB7320
20 stacks PB9320
Power Blotter Pre-cut Membranes and Filters, PVDF, regular size
250 mL PB7100
Power Blotter 1-Step Transfer Buffer (5X) 1L PB7300
Detect Quantity Cat. No.
Bandmate Automated Western Blot Processor 1 system BW1000
24
Multiplex western blotting
Multiplex
western blotting
멀티플렉스 형광 웨스턴 블로팅은 하나의 블랏에서
여러 타겟 단백질을 동시에 분석할 수 있습니다. 이를
통해 시간을 절약하고, 비용을 절감하며, 데이터 생성
및 수집의 효율성을 개선할 수 있습니다. 선명하고
정확한 결과를 얻어내려면 형광 웨스턴 블랏만을
위해 특별히 디자인된 제품을 선택하는 것이
중요합니다.
샘플 준비부터 데이터 수집에 이르는 멀티플렉스
웨스턴 블랏을 위한 전반적인 제품 솔루션에
대해 알아보세요. 자사의 제품들은 정확한 정량
결과와 안정적인 시그널을 제공하여 각 단계를
최적화하고 실패를 방지하며 최상의 결과를
얻을 수 있도록 도와드립니다.
25
Separate Transfer Detect
Fluorescence-
compatible buffer
Sample buffers
containing
bromophenol blue
680 nm
26
Separate Transfer Detect
~70 80
95
26
Visible light
Visible light
15 ~15 15
11 ~10
10
27
Separate Transfer Detect
Nitrocellulose, 0.45 µm
Low-fluorescence PVDF,
0.2 µm
PVDF, 0.45 µm
28
Separate Transfer Detect
EGFR
Stat3
Ras10
29
Separate Transfer Detect
Fluorescent-labeled antibodies
A431 Cas9
EGF A431 control A431 EGFR KO
control
(200 ng/mL, 10 min)
1 2 1 2 1 2 Overlay:
p-EGFR
160 kDa
EGFR
Overlay:
45 kDa p-MEK1 MEK1
37 kDa GAPDH
45 kDa p-MEK1
Fluorescent western blot analysis of p-EGFR, EGFR, p-MEK1, MEK1, and GAPDH in A431 control, Cas9, and EGFR KO cells. Targets
were detected using Alexa Fluor Plus donkey anti-rabbit 680 and 800 conjugates and Alexa Fluor Plus donkey anti-goat 488 and 555 conjugated
secondary antibodies (Cat. No. A32814 and A32816).
30
Separate Transfer Detect
정확한 Total protein normalization 결과를 얻으세요. No-Stain blot image β-Actin
50 40 30 20 10 50 40 30 20 10
웨스턴 블랏을 할 때 Housekeeping 단백질을 사용하면 실험
조건에 따라 발현의 정도가 달라지기도 하고, 때로는 과포화
(oversaturated) 웨스턴 블랏 시그널을 보여 정확한 정량
분석을 방해하기도 합니다. Invitrogen™ No-Stain™ Protein
GAPDH
Labeling Reagent를 사용하면 housekeeping 단백질의 50 40 30 20 10
검출이 필요 없어 housekeeping 단백질이 가져다주는
변동성과 부정확성이라는 두 가지의 단점을 보완할 수 있습니다.
31
Separate Transfer Detect
iBright FL1500 모델은 최대 4개의 형광 웨스턴 블랏 필터 세트는 iBright FL1500 이미징 시스템에 장착되어 있어
멀티플렉싱이 가능한 5개의 형광 채널을 갖추고 있어 하나의 RGB와 NIR 형광 블랏에 활용할 수 있습니다.
블랏에서 여러 단백질을 연구할 수 있습니다. 유의미하고
Example compatible Excitation Filter range Emission Filter range
대표적인 비교를 통해 실험을 향상시킬 수 있습니다. Smart fluorophores channel (nm) channel (nm)
Exposure™ 기술이 탑재되어 각 형광 채널에 대해 개별적으로 Alexa Fluor Plus 488,
EX1 455–485 EM1 508–557
Alexa Fluor 488
신호 대 잡음비(signal-to-noise ratio)가 최적화되므로
Alexa Fluor Plus 555,
EX2 515–545 EM2 568–617
멀티플렉스 형광 웨스턴 블랏 데이터를 더욱 쉽게 얻으실 수 Alexa Fluor 546
Alexa Fluor Plus 647,
있습니다. Alexa Fluor 594
EX3 608–632 EM3 675–720
Four-color fluorescence blot Alexa Fluor Plus 488 dye—RB1 Alexa Fluor 546 dye—calreticulin
protein with GST and HA tag protein (55 kDa)
(134 kDa)
Alexa Fluor Plus 680 dye—p23 Alexa Fluor Plus 800 dye—HSP90 Membrane—iBright prestained marker
protein (23 kDa) protein (90 kDa)
Four-color multiplexed fluorescent blot: false color composite top left, and individual channels shown in grayscale as pictured.
32
Ordering information
Separate Quantity Cat. No.
Fluorescent Compatible Sample Buffer (4X, nonreducing) 5 mL LC2570
iBright Prestained Protein Ladder 2 x 250 µL LC5615
PageRuler Plus Prestained Protein Ladder, 10 to 250 kDa 2 x 250 µL 26619
PageRuler Prestained NIR Protein Ladder 2 x 250 µL 26635
Transfer Quantity Cat. No.
Nitrocellulose/Filter Pre-cut Blotting Membranes, 0.2 µm pore size, (8.3 x 7.3 cm) 20 pack LC2000
Nitrocellulose/Filter Pre-cut Blotting Membranes, 0.45 µm pore size, (8.3 x 7.3 cm) 20 pack LC2001
Nitrocellulose/Filter Pre-cut Blotting Membranes, 0.2 µm pore size, (8.5 x 13 cm) 16 pack LC2009
Nitrocellulose/Filter Pre-cut Blotting Membranes, 0.45 µm pore size, (8.5 x 13 cm) 16 pack LC2006
Nitrocellulose Membrane Roll, 0.45 µm pore size (30 cm x 3.5 m) 1 roll 88018
Low-Fluorescence PVDF Transfer Membranes, 0.2 µm pore size, (7 x 8.4 cm) 10 sheets 22860
Detect Quantity Cat. No.
Blocker FL Fluorescent Blocking Buffer (10X) 100 mL 37565
No-Stain Protein Labeling Reagent 40 rxn A44449
No-Stain Protein Labeling Reagent, Trial Size 10 rxn A44717
iBright FL1500 Imaging System 1 system A44115
33
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34
Optimizing your western blots
Optimizing your
western blots
실험이 항상 성공하지는 않죠—
저희도 그것을 잘 알고 있습니다.
자사의 웨스턴 블로팅 제품들을 사용해서
논문 게재 수준의 블랏 데이터를 얻고 웨스턴
블랏의 효율성을 높여보세요. 자사는 웨스턴
블랏을 최적화 할 제품 및 솔루션과 더불어
실험 팁도 제공하고 있습니다.
35
Separate Transfer Detect
100% 누수 방지 시스템으로 더이상 Gel을 낭비하지 마세요.* SureCast Gel Handcast System의 장점:
Invitrogen™ SureCast™ Gel Handcast System을 통해 • 누수 방지 디자인—Gel을 다루는 시간이 절약되어 시간 활용이
Polyacrylamide mini gel을 간편하게 만들어보세요. SureCast 용이합니다.
system은 자사의 Mini Gel Tank와 완벽하게 호환됩니다.
• 유리 플레이트의 뛰어난 내구성—그 어떤 제조사의 플레이트보다
Invitrogen™ SureCast™ handcasting reagent는 물론,
최대 20배 이상의 견고함을 제공합니다.
일반적으로 사용하는 Polyacrylamide gel 캐스팅 시약을
사용하셔도 됩니다. • 유니크하게 기울어진 디자인—특별 제작된 디자인으로
아크릴아미드 솔루션의 손실을 최소화합니다.
* Terms and conditions apply. For complete details, go to thermofisher.com/surecastterms. • 간단한 조립식의 캐스팅 구성품—하나의 동작만으로도 사용할 수
있으며, load-and-lock 메커니즘을 기반으로 합니다.
Welcome packs
SureCast Gel Handcast Bundle A SureCast Gel Handcast Bundle B
The Invitrogen™ SureCast™ Gel Handcast Bundle A includes all the The Invitrogen™ SureCast™ Gel Handcast Bundle B includes all the
hardware and reagents to get started: hardware to get started:
2 SureCast Gel Handcast Stations 2 SureCast Gel Handcast Stations
2 SureCast Glass Plate Sets (2 front, 2 back) 2 SureCast Glass Plate Sets (2 front, 2 back)
2 SureCast Sealing Pads (attached to handcast stations) 2 SureCast Sealing Pads (attached to handcast stations)
10 SureCast Gel Spacers 10 SureCast Gel Spacers
1 10-Well Multi-Use Tool 1 10-Well Multi-Use Tool
1 12-Well Multi-Use Tool 1 12-Well Multi-Use Tool
1 15-Well Multi-Use Tool 1 15-Well Multi-Use Tool
6 Gel Combs (2 each: 10-, 12-, 15-well) 6 Gel Combs (2 each: 10-, 12-, 15-well)
1 SureCast Stacking Buffer 2-pack
1 SureCast Resolving Buffer 2-pack
1 SureCast APS (25 g)
1 SureCast TEMED (30 mL)
1 SureCast Acrylamide Solution, 40% (450 mL) thermofisher.com/surecast에서 더 자세한 정보를 확인해보세요
36
Separate Transfer Detect
thermofisher.com/proteingels에서 Protein Gel Selection Guide를 이용해 알맞은 protein gel을 찾아보세요
37
Separate Transfer Detect
38
Separate Transfer Detect
Laemmli 시스템(Tris-glycine)은 SDS-PAGE를 사용해 다양한 기존의 Tris-glycine gel과 달리, NuPAGE와 Bolt gels 은
단백질을 분리하는 데 가장 많이 사용되는 Gel 시스템 입니다. 높은 Bis-Tris HCI(pH 6.4) 버퍼 성분으로 약 7.0의 pH를 띕니다.
알칼리성의 pH를 가진 Laemmli 시스템은 단백질이 분해될 수 있는 Bis-Tris 시스템의 중성 pH는 Laemmli 시스템에 비해 다음과 같은
환경으로 밴드 변형, 해상도 저하 등 인공적인 밴드를 나타낼 수 장점을 제공합니다.
있습니다.
• Gel 안정성이 향상되어 8~16개월로 더 긴 저장 수명을 가집니다.
Laemmli 시스템에서 밴드 해상도가 저하되는 주요 원인은 다음과 • 중성 pH에서 전기영동 시 단백질 안정성이 향상되어 밴드 해상도가
같습니다. 더 깨끗하며 정확한 결과를 얻을 수 있습니다.
• Gel의 pH가 높아짐에 따라 단백질의 탈아미노화(Deamination) • Invitrogen™ NuPAGE™ 및 Bolt™ Antioxidant를 사용하면
와 알킬화(Alkylation)같은 화학적 변화가 일어납니다. Reduced protein의 상태를 전기영동 및 단백질의 블로팅 중에
유지할 수 있습니다.
• Gel의 불안정한 산화환원 상태가 Cysteine 함유 단백질에서
Reduced disulfide의 Reoxidation반응이 일어납니다.
39
Separate Transfer Detect
thermofisher.com/wettransfer에서
더 자세한 내용을 확인해보세요
40
Separate Transfer Detect
Blocking buffers
단백질을 연구하기 전 항체의 비특이한 결합을 방지하려면 멤브레인 표면을 Block 해야 합니다.
그렇지 않았을 때, 항체나 다른 검출 시약들이 멤브레인에 있는 다른 사이트와 결합할 수
있습니다. 하나의 Blocking buffer로 모든 웨스턴 블로팅에 일괄적으로 사용할 수 없습니다.
아래의 표를 참조해 최적의 Blocking buffer를 찾아보세요.
phosphoproteins serum albumin fewer chances of cross-reaction with • Best to use when storing concentrate
assay components than serum or reused antibodies in blocker
milk solutions
• Helps minimize or eliminate cross-
reactivity associated with protein-
based blocking buffers
PBS
... you need a Non-protein • Sample-and-antibody combinations • Use when protein-based
Pierce Protein-Free
™
TBS
protein-free blocking require the elimination of all possible blockers cause high
Blocking Buffer PBST
blocker compound exogenous animal proteins in background
TBST
the assay system to avoid cross-
reaction or quenching of the desired
probe function
41
Separate Transfer Detect
• 뛰어난 민감도—Low-picogram에서High-femtogram 까지
커버하는 민감도
• 안정적인 시약— Working solution은 최대 8시간까지 안정하게 • 탁월한 유지력—가장 일반적으로 사용되는 1:5,000에서 1:10,000
사용할 수 있으며 상온에서 1년까지 보관 가능한 키트 의 2차 항체(1 mg/mL) 희석 범위를 벗어나도 우수한 성능 유지
SuperSignal West
Pico PLUS
Bio-Rad Clarity
SuperSignal West Pico PLUS substrate outperforms Bio-Rad™ Clarity™ substrate. Detection of the indicated targets was performed using 2-fold
serial dilutions of HEK293 or HeLa cell lysates. Chemiluminescent detection of indicated target and substrate comparison was performed, following a
5 min incubation with either SuperSignal West Pico PLUS or Bio-Rad Clarity substrate.
42
Separate Transfer Detect
Low-picogram to
Detection level Mid-femtogram Mid- to low-femtogram Low-femtogram to high-attogram
high-femtogram
Signal duration Up to 24 hours Up to 24 hours Up to 8 hours Up to 6 hours
Antibody 1°: 1:1K–5K 1°: 1:1K–50K 1°: 1:5K–100K 1°: 1:1K–1:5K
dilution 2°: 1:10K–1:100K 2°: 1:50K–1:250K 2°: 1:100K–1:500K 2°: 1:100K–1:250K
Superior sensitivity and
Best for use with imaging Most sensitive substrate for HRP
intensity and longer duration Good sensitivity with
Advantages equipment; long-lasting detection with high signal-to-
than other entry-level ECL optimized conditions
signal duration noise ratio
substrates
Routine western blots;
Target is very low in abundance/
working with new protein Maximum signal duration is Target is low in abundance/
sample is precious; requires
Select when: target when western blotting needed; performing sensitive sample is precious; system
maximum sensitivity with less
conditions are not yet quantitation is optimized
optimization
optimized
1.8
Amersham ECL Select Bio-Rad Clarity
GE Healthcare ECL
1.6
Immobilon Western
1.4
Bio-Rad Clarity
1.2
GE Healthcare ECL
1.0
0.625 μg 0.313 μg 0.156 μg
HeLa lysate protein
43
Separate Transfer Detect
thermofisher.com/antibodies에서
당신의 연구에 맞는 항체를 찾아보세요 HRP Secondary Antibody
* Terms and conditions apply. For complete details, go to thermofisher.com/antibody-performance-guarantee.
te
te
te
Ly te
Ly te
Ly te
Ly te
h
h
as
as
as
as
sa
sa
sa
sa
u
u
는 본래의 항체만 검출해 항체의 H-chain 및 L-chain 프래그먼트로
El
El
El
El
W
W
인한 저해를 최소화함으로써 표적 항원을 정확하고 특이적으로 NFκB and Bax were immunoprecipitated from A549 lysate using
Protein A/G agarose resin and rabbit anti-NFκB (panels A and B) and
검출합니다. rabbit anti-Bax (panels C and D). Panels A and C were detected with
goat anti-rabbit HRP, which masked the targets. Panels B and D were
detected with the Clean-Blot IP Detection Reagent (HRP), unambiguously
revealing the target protein without interference.
44
Separate Transfer Detect
15 min
웨스턴 블랏을 Reprobing하면 시간을 아끼고 샘플을 보존할 수 Thermo Scientific™ Restore™ stripping buffers 는 안전하고
있습니다. 특화된 버퍼는 블랏에서 1차, 2차 항체를 떼어내 다른 효과적으로 항체를 제거해 reprobing할 수 있습니다.
타겟 단백질의 항체를 reprobing할 수 있는 조건을 만들어 줍니다.
45
Ordering information
Separate Quantity Cat. No.
SureCast Gel Handcast Bundle A - Hardware and Reagents 1 kit HC1000SR
SureCast Gel Handcast Bundle B - Hardware Only 1 kit HC1000S
Bolt Bis-Tris 4–12%, 10-well Welcome Pack 1 kit NW0412A
Bolt Bis-Tris 4–12%, 15-well Welcome Pack 1 kit NW0412B
NuPAGE Bis-Tris 4–12%, 10-well Welcome Pack 1 kit NP032A
NuPAGE Bis-Tris 10%, 10-well Welcome Pack 1 kit NP030A
NuPAGE Tris-Acetate 3–8%, 10-well Welcome Pack 1 kit EA0375A
Novex Tris-Glycine 10%, 10-well Welcome Pack 1 kit XP0010A
Novex Tris-Glycine 4–12%, 10-well Welcome Pack 1 kit XP0412A
Novex Tris-Glycine 10%, 15-well Welcome Pack 1 kit XP0010C
Novex Tris-Glycine 4–12%, 15-well Welcome Pack 1 kit XP0412C
Transfer Quantity Cat. No.
Mini Blot Module 1 unit B1000
Surelock Tandem Midi Blot Module 1 unit STM2001
Detect Quantity Cat. No.
StartingBlock (TBS) Blocking Buffer 1L 37542
StartingBlock T20 (TBS) Blocking Buffer 1L 37543
StartingBlock (PBS) Blocking Buffer 1L 37538
StartingBlock T20 (PBS) Blocking Buffer 1L 37539
Blocker FL Fluorescent Blocking Buffer (10X) 100 mL 37565
Pierce Clear Milk Blocking Buffer (10X) 100 mL 37587
Blocker BSA (10X) in TBS 125 mL 37520
Blocker BSA (10X) in PBS 200 mL 37525
Pierce Protein-Free (TBS) Blocking Buffer 1L 37570
Pierce Protein-Free T20 (TBS) Blocking Buffer 1L 37571
Pierce Protein-Free (PBS) Blocking Buffer 1L 37572
Pierce Protein-Free T20 (PBS) Blocking Buffer 1L 37573
Pierce ECL Western Blotting Substrate 250 mL 32209
Pierce ECL Plus Western Blotting Substrate 100 mL 32132
SuperSignal West Pico PLUS Chemiluminescent Substrate 200 mL 34577
SuperSignal West Dura Extended Duration Substrate 100 mL 34075
SuperSignal West Femto Maximum Sensitivity Substrate 100 mL 34095
SuperSignal West Atto Ultimate Sensitivity Substrate 100 mL A38555
46
Get the high-quality western blots you need with educational and product
support at your fingertips
47
thermofisher.com/western에서
웨스턴 블랏 제품의 더 많은 정보를 확인해보세요
www.thermofisher.com
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