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Related Products for Blot Detection

Catalog # Description
Instruction Manual
161-0385 Precision Plus Protein WesternC Pack, (Precision Plus
Protein WesternC standards, StrepTactin-HRP
Conjugate, 0.125 ml)
161-0380 Precision Protein StrepTactin-HRP Conjugate, 0.3 ml
Precision Plus
161-0381 Precision Protein StrepTactin-HRP Conjugate, 0.125 ml
170-5070 Immun-Star WesternC™ Chemiluminescent Kit
Protein™
170-5044 Immun-Star GAM-HRP Detection Kit
170-5045 Immun-Star GAR-HRP Detection Kit WesternC™
170-5047 Immun-Star GAM-HRP Conjugate, 2 ml
170-5046 Immun-Star GAR-HRP Conjugate, 2 ml
170-5010 Immun-Star AP GAM-AP Detection Kit
Standards
170-5011 Immun-Star AP GAR-AP Detection Kit
161-0382 Precision Plus StrepTactin-AP Conjugate, 0.3 ml

Catalog #161-0376
Product shipped at 4°C.
Store at –20ºC upon arrival.

Bio-Rad Laboratories, Inc.


2000 Alfred Nobel Dr., Hercules, CA 94547 USA
(510) 741-1000 10008761 Rev B

Precision Plus Protein WesternC standards are


imagers kD
Gel Blot intended for use in a wide range of applications and
A representative lot of
weight on film and CCD 250 allow easy visualization during electrophoresis, blot
10

Precision Plus Protein


Estimating molecular Blot development transfer, and chemiluminescent development of western
15
150
WesternC standards on blots on film or by CCD imagers.
20
efficiency
a Criterion™ 4–20% 100 25
Assessing transfer 75
Blot transfer Application Use
Tris-HCl gel and a 37
western blot detected
electrophoresis Gel electrophoresis Monitoring migration during
by chemiluminescence
50
by chemiluminescence electrophoresis
50
Monitoring migration during Gel electrophoresis western blot detected
37
Tris-HCl gel and a
Use Application Blot transfer 75
Assessing transfer
25 100 a Criterion™ 4–20%
efficiency
blots on film or by CCD imagers. 20 WesternC standards on
150
15
transfer, and chemiluminescent development of western Blot development Estimating molecular
Precision Plus Protein
10
allow easy visualization during electrophoresis, blot 250 weight on film and CCD
A representative lot of
intended for use in a wide range of applications and Blot Gel
kD imagers
Precision Plus Protein WesternC standards are
10008761B.qxp 3/4/2008 10:29 AM Page 3

Product Description 7. Probe with a secondary antibody appropriate


Precision Plus Protein WesternC standards offer 10 to the protein of interest, and include
distinct, prestained protein bands with a molecular approximately 1 µl of StrepTactin-HRP1 or
weight of 10, 15, 20, 25, 37, 50, 75, 100, 150, and StrepTactin-AP per 10 ml of secondary
250 kD. The 25, 50, and 75 kD bands are stained incubation solution to detect the bands of the
pink for easy band referencing and blot orientation. Precision Plus Protein WesternC standards
8. Wash the blot
Precision Plus Protein WesternC standards contain an 9. Develop with a suitable development reagent,
integral Strep-tag sequence, enabling parallel detection such as the Bio-Rad Immun-Star HRP kit or
on western blots using StrepTactin-HRP (catalog Immun-Star AP kit (for film detection), or
#161-0380, 161-0381) or StrepTactin-AP Immun-Star WesternC kit (for CCD imaging)
(catalog #161-0382) and HRP or AP-conjugated
Note: The recommended dilution of the
secondary antibodies specific to the protein of interest.
StrepTactin-HRP and StrepTactin-AP conjugate
The molecular weights of the Precision Plus Protein is 1:10,000–1:50,000 for chemiluminescent
WesternC standards are confirmed by migration in a detection2 with the Immun-Star WesternC kit
Laemmli SDS-PAGE system. The sharp ladder bands (#170-5070 or similar).
provide an accurate estimation of the molecular
weight of unknown proteins, with an R2 value >0.99 Specifications
on a 4–20% Tris-HCl SDS-PAGE gel.* Catalog # 161-0376
Volume 250 µl
Number of 50
Instructions for Use applications
Precision Plus Protein WesternC standards are Loading buffer 30% (w/v) glycerol, 2% SDS,
62.5 mM Tris, pH 6.8, 50 mM DTT,
provided in loading buffer and require no dilution.
5 mM EDTA, 0.02% NaN3
Before use, allow the tube to reach room
Shelf life Minimum 12 months at –20°C; if the
temperature and mix thoroughly. This will ensure that product has been stored as directed
any solids that may have precipitated at –20°C have for longer than 12 months, the shelf
returned to solution. Do not heat the product above life may be extended by adding
room temperature. dithiothreitol (DTT) to approximately
50 mM
For chemiluminescent detection of the Precision
Recommended Loading Volumes
Plus Protein WesternC standards: Load Volume3 Application
1. Load 5 µl of the Precision Plus Protein 5 µl Chemiluminescent blot development
WesternC standard onto the gel next to
protein of interest Strep-tag technology for western blot detection is covered by
2. Run the SDS-PAGE gel at the appropriate US patent 5,506,121 and by UK patent 2,272,698. StrepTactin
is covered by German patent application P 19641876.3. Bio-Rad
voltage, current and time Laboratories, Inc. is licensed by Institut für Bioanalytik GmbH to
3. Transfer the protein to a membrane at the sell these products for research use only.
appropriate voltage and current
4. Block the membrane in blocking solution 1 StrepTactin-HRP and StrepTactin-AP are designed not to
5. Probe with a primary antibody specific to the interfere with secondary antibody incubation.
2 For longer exposure times, use of a greater dilution of
protein of interest
6. Wash the blot StrepTactin-HRP (1:50,000–1:100,000) will prevent
overexposure of the bands of the Precision Plus Protein
WesternC standards.
*Migration pattern of the 25, 50, and 75 kD bands may differ on
3 For very short exposure times, or for enhanced band intensity
neutral pH gels (Criterion XT gels or similar).
on a gel or a blot, an optional load volume of 10 µl may be
used.