PROCAP DYKDDDDK-Agarose IP/CoIP KIT

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China

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(Negotiable) Get Latest Price

Place of Origin:

China

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Delivery Time:

7-15 days

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Product Group :

Protein Biology

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Product Description

PROCAP DYKDDDDK-Agarose IP/CoIP KIT CAT：PFA**5 Price: $**5 for *5T

Catlog No.：PFA**5
Storage conditions:Stored at **0℃for one year, Long-term storage at **0℃to avoid repeated freezing and thawing.

Component:

Cat.No	Name	Specification
FNA****0/FNA*****0	DYKDDDDK-Nanoab-Agarose	*0ul(5T)/**0ul(0T)
IP**1A	Lysate A	*0ml
IP**1B	Lysate B	*0ml
IP**1C	solution C	*0ml
IP**1D	solution D	*0ml
IP**1E	solution E	3x1ml

Product description:
Agarose beads of coupling anti-DYKDDDDK nanoantibody are used to immunize DYKDDDDK fusion protein.
Experimental steps:
Plant tissue cracking treatment:
Take an appropriate amount of plant tissue samples (leaves, etc.) and place them in frozen liquid nitrogen, and then put the frozen plant tissue samples in a mortar for grinding, so as to fully grind and destroy their cell walls. Add *******0ul lysate A or lysate B (added with protease inhibitor PMSF, etc.) for cracking. In order to improve the cracking efficiency, add **0ul glass powder and fully shake for *0min. After cracking, ****0 rpm, centrifuge for *0min, suck the supernatant and place it in a new centrifuge tube, and discard the sediment.
Collect cells
Approximately ******7 cells expressing DYKDDDDK fusion protein were used for each immunoprecipitation reaction, and the number of cells could be appropriately adjusted according to the expression of DYKDDDDK fusion protein. Suck out the medium and add precooled 1 × PBS, rinse twice, collect adherent cells by cell scraping or trypsin digestion, transfer the cells to a centrifuge tube, centrifuge **0g for 3 minutes and discard the supernatant.
Cell lysis
1. Protease inhibitors are added to the Lysate A or Lysate B, and the cells are suspended with a precooled lysis buffer of **0μL.
2. Put it on the ice for *0 minutes and blow it fully every *0 minutes.
3.Centrifuge ****0 g at 4℃for *5 minutes, transfer the cracking product (supernatant) to a new precooling tube, and discard the precipitation.

Note: At this time, the cell lysis product can be preserved at **0°C for a long time.

Balanced beads
4. Fully mix DYKDDDDK-Nanoab-Agarose, absorb *0μL of the product into the **0μL precooled cracking buffer, 4℃ 2,**0g centrifugal for 2 minutes, and discard the liquid. (This step is optional)
Binding protein
5. The balanced DYKDDDDK-Nanoab-Agarose Beads are added to the cell lysis product (*0μL can be added directly to the cell lysis product if step 4 is not done) and rotate and combine at 4°C for 5 to *0 minutes. The combination time can be adjusted according to the needs of the experiment. If necessary, the *0μL cracking product is retained for immunoprinting analysis.
6. 4℃, ***0g centrifugal for 2 minutes, discard the liquid. If necessary, *0μL supernatant is retained for immunoprinting analysis.
Wash the beads
7. Use **0 μL precooled pyrolysis buffer resuspended DYKDDDDK-Nanoab-Agarose, centrifugation for 2 minutes under 4℃, ***0g, discarded the supernatant and washed again for 2 times. Minimize cleaning time.
Elution protein
Method 1:
8. Adding *0μl 2×SDS-sample buffer suspends DYKDDDDK-Nanoab-Agarose.*5℃, heated for *0 min, fully denatured, centrifugation for 2 minutes under 4℃, ***0g, collected the supernatant and the collected products can be analyzed by SDS-PAGE and immunoblotting
Method 2:
9. Alternative Step 8: Add *0μL Elution E to elute the bound protein for a *0 second incubation period, ***0g centrifugal 2 minutes to collect and clear, and immediately add 5μL 1.0 M Tris (pH*0.4) to neutralize the acidic glycine.
Note: This step can be repeated to improve the efficiency of elution.

Matters needing attention:
1. Selection of cracking fluid and cracking fluid:
Lysate A is a mild lysate, and lysate B is a strong lysate. Please select the corresponding lysate according to your own experimental samples, for example, if it is cytoplasmic protein, you can choose lysate A or mix A with B, if it is nuclear protein, you can choose lysate B.
Selection of rinse solution C and rinse solution D: (lysate D is a high-salt solution, which may destroy the weak interaction between proteins), if the interaction between two proteins is strong, but also want to reduce non-specific binding, choose D; If the interaction between two proteins is weak, C is preferred.
2. In order to achieve the best use effect, try to avoid excessive repeated freezing and thawing. It can be used after proper sub-packaging.
3. This product is only used for scientific research by professionals, not for clinical diagnosis or treatment, nor for food or medicine

Country:	China
Model No:	-
FOB Price:	(Negotiable) Get Latest Price
Place of Origin:	China
Price for Minimum Order:	-
Minimum Order Quantity:	-
Packaging Detail:	-
Delivery Time:	7-15 days
Supplying Ability:	-
Payment Type:	T/T, L/C, D/A, D/P, Western Union, Money Gram, PayPal, Other
Product Group :	Protein Biology