SNAP i.d. Workflow Value Proposition

1.

2. SNAP i.d. Workflow Value Proposition

Western Blotting Protocol
Sample
Prep
45 min
-2 Hrs
Electrophoresis
Membrane
Transfer
Blocking
Antibody
Addition
Detection
.5 -1 Hr
1-2.5 Hrs
1 Hr
3 Hrs
15 min
SNAP i.d.™
Higher quality blots with Immunodetection occurring
in 30 min vs. 4 hrs
• Reduces incubation times in Western Blotting
• Compatible with all reagents and membranes
• No directly competitive product on the market

3. SNAP i.d. Workflow Value Proposition

Western Blotting Protocol
Sample
Prep
Electrophoresis
Membrane
Transfer
Blocking
Antibody
Addition
Detection
SNAP i.d.™
Standard
Higher quality blots
in less than 30 min
5%NFDM
0.05% NFDM

4.

Product Contents

5. SNAP i.d. Components

A.
B.
C.
D.
E.
7.5 cm x 8.8 cm
A
4.6 cm x 8.8 cm
SNAP i.d. Base
Single Blot Holder, 30/pk
Double Blot Holder, 30/pk
Triple Blot Holder, 20/pk
Antibody Collection Tray,
20/pk
B
3.2 cm x 8.8 cm
C
D
E

6. Additional Items

Line Filter Millex-FA50
(SLFA05010)
Perforated Stopper
(XX1004708)
Vacuum source
must provide
4” Hg (135 millibar)
Silicone Tubing
(XX7100004)
1 Liter Vacuum Flask
(XX1004705)
Chemical duty pump
(WP6111560, WP6122050)

7. SNAP i.d. SKU’s

Item Description
WBAVDBASE
SNAP i.d. Protein
Detection System
WBAVDBH01
Single Blot Holders
including Spacers, 30/pk
WBAVDBH02
Double Blot Holders
including Spacers, 30/pk
WBAVDBH03
Triple Well Blot Holders
including Spacers, 20/pk
WBAVDABTR
Antibody Collection Tray,
20/pk
WBAVDR0LL
Blot Roller, 1/pk
“zero”
Hardware
Consumables
SKU
Internal Project
Codename
Western Blotting AVIDXXXX

8. SNAP i.d. Protocol Overview: Blot Holder Assembly

1) Open Blot Holder
2) Wet Flow Distributor
3) Place Blotted Membrane
4) Place Spacer
5) Close Blot Holder
6) Turn over Blot Holder

9. SNAP i.d. Protocol Overview: Immunodetection

1) Open Lid
2) Insert Blot Holder
3a) Add Blocking Solution
3b) Turn on Vacuum
4) Turn off Vacuum
5a) Add Primary Antibody
5b) Incubate 10 min
5c) Turn on Vacuum
6a) Add wash solution (3x)
6b) Turn off Vacuum
Repeat steps 4-6 for 2 Ab

10. Feature and Benefits Summary

Features
Benefits
Actively driven reagent flow with vacuum
Minimal dead volume
Reduction of Immunodetection time from 4
hours to 30 min
Integrated flow distributor in Blot Holders
Ensures consistent results equivalent to
standard WB protocol
Blot holders accept: 7.5 x 8.8 cm
4.6 x 8.8 cm
3.2 x 8.8 cm
System compatible with common blot sizes
(80% of market)
2 blot holders can be run independently or in
parallel
Up to 6 blots can be processed
simultaneously
Validated with chemiluminescent and
fluorescence detection
Compatible with existing Western Blotting
protocols
Antibody collection tray
Allows for collection of primary antibodies for
reuse
Integrated vacuum regulator
Ensures consistent vacuum pressure without
the need for an external regulator

11. Product Position

• Superior Blots in a SNAP!
– Optimize your blots
– Increase the quality of your blots
– And do it FAST!
– Results for your afternoon meeting! (Customer quote)

12. Product position: Optimization

• All blots should be optimized but most researchers don’t have the time to
do this tedious process.
• SNAP i.d. makes optimization easy by allowing 6 blots to be processed
simultaneously in only 30 minutes!
• What do they typically optimize?
–
–
–
–
Antibody concentrations/dilutions
Detection reagent conditions
Target protein concentrations (more rarely)
Blocking conditions (a little secret to sensitivity)
• SNAP i.d. permits fast optimization for better results.
• Point out the Immunodetection handbook Western blotting section for
helpful hints.

13. How it Works

• Traditional western blotting takes a variety of formats and reagent
conditions to accomplish. It’s a passive process!
• SNAP i.d. actively drives reagents through the membrane to increase
the quality of the blots and increase the speed of immunodetection!
• It’s a combination of reagent flows and concentrations
Vs.
Standard ‘rocking’ of
reagents
Actively drive reagents with
vacuum flow

14. How it Works – reagent flows

Reagents penetrate more of the membrane 3D structure
where the proteins are blotted.
Result = Increase quality of the blot in a SNAP!
Standard
Gentle Rocking
Reagents diffuse
slowly into membrane
Reagents rapidly
driven into membrane
Vacuum

15. How it Works – reagent flows

Blocking
• Efficient coverage of membrane which yields higher sensitivity
– Can use 1/10th-1/100th less concentrated blocking solution to minimize
overblocking
• Actively driven vacuum flow coats inner surfaces of membrane in 20 sec
Standard
GAPDH
1
2
3
4
5
6
5%NFDM
7
8
1
2
3
4
5
6
0.5% NFDM
7
8
1
2
3
4 5
6
7
8
0.1% NFDM
1
2
3
4
5
6
7
0.05% NFDM
8

16. Compatible Blocking Reagents and Recommended Concentrations

How it Works – reagent flows
Washing
• Unbound Antibodies are thoroughly flushed out of the
membrane
– Yields lower backgrounds which may yield higher Sensitivity
Standard
GAPDH
1
2
3
4
5
6
5%NFDM
7
8
1
2
3
4
5
6
0.5% NFDM
7
8
1
2
3
4 5
6
7
8
0.1% NFDM
1
2
3
4
5
6
7
0.05% NFDM
8

17. How it Works – reagent flows

Standard vs. SNAP i.d. - concentrations
Concentrations
• Blocking concentrations are limited to prevent clogging of blot holder
• Antibody concentrations are increased to speed up reaction kinetics
Step
Standard Protocol
SNAP i.d.
5% NFDM
0.5% NFDM
Primary Antibody
1X
3X in 1/3 volume
(same quantity)
Washing (3x)
1X
1X
Secondary Antibody
1X
3X in 1/3 volume
(same quantity)
Washing (3x)
1X
1X
Blocking

18. Standard vs. SNAP i.d. - concentrations

Concentrations: The Rule of 3.5
Rule of 3.5
1° & 2° Ab concentration
and volume
Blocking concentration
Use 1/3 the volume of Ab at 3x concentration
Concentration: Drives Ab incubation kinetics faster
Dead volume: Minimizes Ab consumption
Block in 0.5% NFDM vs 1.0%-5.0% NFDM
To prevent clogging of flow distributor
To minimize overblocking and maximize sensitivity

19. Concentrations: The Rule of 3.5

How it Works: Time savings
Western Blotting Protocol
SNAP i.d.™
Sample
Prep
Standard
Electrophoresis
Membrane
Transfer
Blocking
1° Antibody
Addition &
Incubation
1 Hr
1 Hr-overnight
Blocking
Antibody
Addition
Detection
Washing
2° Antibody
Addition &
Incubation
Washing
15 min
1 Hr
15 min
4 Hrs
Vs.
20 sec
10 min
1 min
10 min
1 min
22 min

20. How it Works: Time savings

SPIN Analysis
Situation
Problem
Implication
Need Payoff
Home-Made
Protocol
Takes a long time
- Can only do one
a day
- Results aren’t
optimized so
quality is variable
- Time wasted
- Not the best
quality
Quality
improvement
FAST!