This product demonstrates separation ability and band sharpness comparable to conventional products, despite a short running time of about 10 minutes.

Product Name	Bullet PAGE Plus	Bullet PAGE One	Company A	Company B
Gel %	5 - 20%	5 - 20%	4 - 20%	4 - 12%
Product No.	21794-54	13064-64	-	-
Electrophoresis Time	10 min 07 sec.	35 min 12 sec.	29 min 35 sec.	48 min 32 sec
Electrophoresis Images

Electrophoresis condition

Bullet PAGE Plus Precast Gel shows a transfer efficiency equialent to company A products.

Transfer solution	Bullet Semi-dry Transfer one (#15353-01)
Transfer condition	25V, 10 min
Gel	Bullet Page Plus 5-20%	Company A 4-20%
Image of after transfer

Sample
① Protein Ladder One Plus (Triple-color) (#19593-25) Dilute Protein Market (10x) (#29458-24) to achieve
② Protein Markers(3x) 2 µl added
➂ Protein Markers(1x) 2 µl added
④Protein Markers(1/3x) 2 µl added

PVDF membrane staining
Stain PVDF membrane with CBB Stain One (ready to use) (#04543).

 Western blotting: Detection of proteins by different sizes

Three proteins of different sizes were detected by western blotting after running by using this product. A wide range of proteins of different molecular weights, from low (Histone H3) to high (EGFR), could be detected, indicating that the product is applicable to western blotting.

Protein detection	EGFR (175 kDa)	β-Actin（42 kDa）	Histon H3 (17 kDa)
Blotting image

(Condition)

Sample:

HeLa cell suspension (10 µg to 2-fold dilution series)

SDS-PAGE:

Bullet PAGE Plus Precast Gel, 5-20%, 17wells (#21794-54) 400 V

Transfer:

Bullet Semi-dry Transfer One (#15353-01) 25 V 10 min. Use PVDF (bore size; 0.45µm)

Blotting:

Bullet Blocking One for Western Blotting (#13779) for 5 mins at RT

Primary antibody:

[EGFR]	EGF Receptor (D38B1) XP Rabbit mAb (CST #4267) 2,000x dilution Signal Enhancer HIKARI for Western Blotting and ELISA Dilute by Signal Enhancer HIKARI for Western Blotting and ELISA (#02270-81) A solution for 60 mins at RT
[β-Actin]	β-Actin antibody(C4) (Santa Cruz #sc-47778) 1,000 x dilution Dilute by Bullet ImmunoReaction Buffer (#18439-85) for 30 mins at RT
[Histone H3]	Histone H3 (D1H2) XP Rabbit mAb (CST #4499) 2,000x dilution Dilute by Bullet Blocking One for Western Blotting (#13779) for 60 mins at RT

Secondary Antibody：

[EGFR]	Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP](Pre-adsorbed) (Novus #NB7187) 50,000x dilution Signal Enhancer HIKARI for Western Blotting and ELISA Dilute by Signal Enhancer HIKARI for Western Blotting and ELISA (#02270-81) B solution for 60 mins at RT
[β-Actin]	β -Actin HRP-labeled Anti-Mouse IgG (self-prepared) 5,000x dilution Diluted by Bullet ImmunoReaction Buffer (#18439-85) for 30 minutes at RT
[Histone H3]	Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) (Novus #NB7187) 30,000x dilution Diluted by Bullet Blocking One for Western Blotting (#13779) for 60 minutes at RT

Detection:

[EGFR] [β-Actin] [Histone H3]

Chemi-Lumi One Super (#02230) Chemi-Lumi One L (#07880) Chemi-Lumi One Super (#02230)

ChemiDoc Touch MP 3 × 3 binning (Bio-Rad Laboratories) (AUTO mode)

The following types of gradient and homogeneous gels are available.

Gel		Gradient Gel
Gel concentration		5 - 10%	5 - 15%	5 - 20%	7.5 - 15%	10 - 20%
Product number	13 wells	21789-34	21791-84	21793-64	21795-44	21797-24
	17 wells	21790-94	21792-74	21794-54	21796-34	21798-14
Images of protein (SDS-PAGE)
Images of nucleic acids (PAGE)

Gel		Homogeneous Gel
Gel concentration		7.5 %	10 %	12.5 %	15 %
Product number	13 wells	21799-04	21801-44	21807-84	21853-74
	17 wells	21800-54	21806-94	21811-14	21854-64
Images of protein (SDS-PAGE)
Images of nucleic acids (PAGE)

If use a power supply with maximum 300 V, please refer to the following running times. The running time is the time it takes for the BPB in the sample buffer to migrate to 5 mm above the lower edge of the gel. The running time varies slightly depending on the temperature of the running buffer and the gel concentration.

For protein analysis (SDS-PAGE), use 25 mM Tris, 192 mM glycine and 0.1% SDS buffer. For nucleic acid analysis (PAGE), use 25 mM Tris, 192 mM glycine buffer. It is recommended that no more than half the well volume (20 µL for 13 wells and 14 µL for 17 wells) of adding samples.

<Notice>

• Use tank which can fit plates of size W100 mm × H80 mm × T3.2 mm.
  Recommended model: WEP-MN Vertical Electrophoresis Tank (Gellex International #WEP-MN)
• Check the suitability of the electrophoresis system (e.g. plate size) and the performance of the power supply (e.g. power range) before use.

Instruction: Bullet PAGE Plus Precast Gel

1. Asano, Kosuke, et al. "An insulin-inducible transcription factor, SHARP-1, represses transcription of the SIRT1 longevity gene." Biochemistry and Biophysics Reports 22 (2020): 100743.