Chemi-Lumi One Markers Kit_{for SDS-PAGE and Western Blotting}

Contains 8 biotinylated proteins as molecular weight markers (M.W. 6,500 - 200,000 Da)
Includes HRP conjugated Strepavidin to detect biotinylated proteins

Western Blotting for detecting of β-Actin

➀Chemi-Lumi One Markers
➁Detection of β-Actin from Hela cell protein extraction solution

Electrophoresis	SDS-PAGE (4-20% gradient gel) Chemi-Lumi One Markers (5 μl)
Membrane:	PVDF	10V, 30 min
Blocking	Blocking One (Product No. 03953-95)	60 min
Primary Ab	Mouse anti-β-Actin, mono (200x)	60 min
Secondary Ab	Goat anti-mouse IgG, HRP (5000x) + Streptavidin (HRP conjugate)(10,000x)	60 min
Detection	Chemi-Lumi One Super (Product No. 00230-30) Chemi-Lumi One L (Product No. 07880-70)

PKG Size: 50 μl
Solution of 8 biotinylated proteins

PKG Size: 250 μl
Horseradish peroxidase conjugated streptavidin solution

Note: The molecular weight of Chemi-Lumi One Markers may slightly differ from unmodified proteins because of biotinylation.

Thaw Chemi-Lumi One Markers and mix thoroughly prior to use.
Add 4 μl of deionized water, 5 μl of Sample Buffer Solution with 2-ME (2x) (Product No.30566) and 1 μl of Chemi-Lumi One Markers to a plastic tube (0.5-1.5 ml volume size). Mix thoroughly.
The mix ratio of deionized water: Sample Buffer Solution with 2-ME (2x) : Chemi-Lumi One Markers is 4: 5 :1.
Cover the tube and incubate in a water bath (approx. 90oC) for 4 minutes.
Cool the tube at room temperature for 10 minutes.
Take 2-10 μl of the diluted Chemi-Lumi One Markers by micropipette and apply carefully into a well of a gel.
Perform SDS-PAGE, western blotting and reaction of primary antibody.

Thaw Streptavidin (HRP conjugate) and mix gently prior to use.
In case of 10cm x 10cm membrane, add 5 μl of Streptavidin (HRP Conjugate) in 50 ml of solution of HRP-conjugated secondary antibody. Mix gently. The mix ratio of Streptavidin (HRP Conjugate): secondary antibody solution is 1: 10,000.
Cover the membrane with above solution [Streptavidin (HRP Conjugate) - secondary antibody mixed solution] and let stand at room temperature for 1hour.
Wash the membrane with 100 ml tPBS (PBS including 0.05% Tween®20) for 3minutes, and then dispose tPBS . Repeat wash three times.
Detect proteins on membrane with one of the following detection reagents.
- Chemi-Lumi One L (Product No.07880)
- Chemi-Lumi One Super (Product No.02230)
- Peroxidase Stain Kit (Product No.26652)

Product Name	Grade	Storage	Catalog Number	PKG Size	Price
Chemi-Lumi One Markers Kit	SP	-20°C	06456-70	1 KIT

Product Name	Grade	Storage	Catalog Number	PKG Size
Gel Casting Reagents
30(w/v)% - Acrylamide/Bis Mixed Solution (37.5:1)	SP	R	06144-05	500 ml
30(w/v)% ‐ Acrylamide/Bis Mixed Solution (29:1)	SP	R	06141-35	500 ml
30(w/v)% ‐ Acrylamide/Bis Mixed Solution(19:1), Nuclease tested	SP	R	07175-75	500 ml
40(w/v)% ‐ Acrylamide/Bis Mixed Solution (37.5:1)	SP	R	06121-95	500 ml
40(w/v)% ‐ Acrylamide/Bis Mixed Solution (29:1)	SP	R	06119-45	500 ml
40(w/v)% ‐ Acrylamide/Bis Mixed Solution(19:1) , Nuclease tested	SP	R	06140-45	500 ml
Separating Gel Buffer Solution(4x) for SDS-PAGE	SP	RT	30651-05	500 ml
Stacking Gel Buffer Solution(4x) with Dye for SDS-PAGE	SP	R	09268-34	100 ml
Stacking Gel Buffer Solution(4x) for SDS-PAGE	SP	R	09267-44	100 ml
Stacking Gel Buffer Solution(4x) for SDS-PAGE	SP	R	32158-25	500 ml
10(w/v)%-Ammonium Peroxodisulfate Solution	SP	F	02634-34	10 ml
N,N,N',N'-Tetramethylethylenediamine	SP	RT	33401-72	25 g
Running Buffer
Running Buffer Solution(10x) for PAGE	SP	RT	30340-91	1 L
Running Buffer Solution(10x) for SDS-PAGE, Tris-Glycine	SP	RT	30329-61	1 L
Gel Steins
CBB Stain One(Ready To Use)	SP	RT	04543-51	1 L
Rapid Stain CBB Kit	SP	RT	30035-14	1 set
Dispotray-SS	-	RT	19386-74	40 pieces
Dispotray-S	-	RT	16526-82	25 pieces
Dispotray-M	-	RT	16551-84	20 pieces
Western Blotting
Chemi-Lumi One L	SP	R	07880-70	1 KIT
Chemi-Lumi One Super	SP	R	02230-30	1 KIT
Peroxidase Stain Kit for Immuno-blotting, Nuclease tested	SP	F	26652-70	1 KIT
Blocking One	SP	R	03953-95	500 ml
Blocking One-P	SP	R	05999-84	200 ml