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Four new products join forces, Abbkine gives you unlimited possibilities

Date:2021-06-28 Views:164

Remember in April, Abbkine solemnly launched 4 high-traffic products? Universal internal reference antibody cocktail set, antibody diluent, enhanced/hypersensitive ECL luminescent solution! These 4 products are a set of meticulous and brand-new products launched by Abbkine after months of painstaking research. Once launched, they have received unanimous praise from customers. In order to help everyone better understand the product information, the data of the 4 products are summarized now, let’s take a closer look:

KTD101-EN Universal Internal Reference Cocktail

Application principle: internal reference of WB experiment needs to consider at least two factors: experimental system and sample to be tested. Due to its relative stability and broad spectrum, internal reference proteins are widely used as internal reference for WB experimental system, and can be used as a semi-quantitative analysis standard. On the other hand, there are some uncertainties in the extraction and preparation of samples to be tested, so it is necessary to select the appropriate total protein as the positive control of WB samples to be tested. The Universal Reference Antibody Cocktail Kit is designed specifically for WB experiments to provide a real reference solution.
The cocktail set contains:

①Classical GAPDH, β-actin and β-tubulin, the three most commonly used reference antibodies cited in hundreds of literatures, can be used in human, mouse and rat samples, which are widely used in internal reference control of different sample types and molecular weights.
②Even if the WB positive control is used, the sample can be directly injected to provide a real reference for WB experiment;
③Superkine ™ Enhanced Antibody Diluent enhances WB target band signal and eliminates background interference, making WB experiments easier. Cocktail sets are optimized for optimal use. The corresponding products in the package (A01020, A01010, A01030, BMU103-CN) can also be ordered separately.

Article Number Cocktail Component Size Application Specification Preservation Condition
A01020-SK GAPDH mouse monoclonal antibody(2B5) 60 μL*1tube The target band is 37 kD and is widely used -20℃
A01010-SK β-Actin mouse monoclonal antibody(1C7) 60 μL*1tube The target band is 43 kD and is widely used -20℃

 

A01030-SK β-Tubulin mouse monoclonal antibody(3G6) 60 μL*1tube The target band was 55 kD -20℃

 

BMP1002-SK WBpositive control 100 μL*1tube The WB positive sample was used as control -20℃
BMU103-SK SuperKine™ Enhanced antibody diluent 50 mL*2bottle Ready-to-use reagent that boosts signal and lowers background 4℃

 

 

Application kit:

a. It is recommended that the experimental dilution ratio of antibody WB is 1:1000-1:10000 (recommended 1:2000). The selection of reference antibodies should ensure that the molecular weight difference between the target protein and the reference protein is more than 5 kD, and the reference protein in the sample is highly expressed and not affected by experimental variables. Contains stabilizer, preservative and glycerin. It is recommended to store at -20℃ for long-term storage after packing.
b. WB positive control is ready-use reagent, which can be directly used for SDS-PAGE loading. The recommended loading volume is 5-10μl, which can be used as positive control in WB experiment. Contains glycerin. It is recommended to store it at -20℃ for long-term storage.
c. Superkine ™ Enhanced Antibody Diluent is a ready-to-use reagent that can be used directly in the dilution and preparation of various primary or secondary antibodies to enhance Wb target protein signaling and reduce background. Diluted antibodies should be stored at 4℃ immediately after antibody incubation to facilitate subsequent reuse.

BMU101-EN/BMU102-EN enhanced/ultra-sensitive ECL luminescent liquid

Application principle: chemiluminescence technology after years of development, there have been a variety of luminous systems, but the most commonly used laboratory is still based on Luminol or its derivatives (isoluminol, etc.) technology system. Western fluorescent ECL immune molecularly imprinted chemiluminescence detection reagent solution is a kind of aims to non radioactive glow (horseradish peroxidase) system, fixed detection in solid phase membrane (such as NC, PVDF) trace protein, the X-ray radiation, the film sensitive or chemiluminescence imaging recording its auxiliary reagents Western blot experiments.

Application Suggestions:

a. Routine electrophoresis, membrane transfer, HRP labeled antibody incubation and membrane washing. Abbkine Enhanced Antibody Diluent (BMU103-CN) is recommended for antibody dilution. At the same time, the universal reference antibody cocktail set (KTD101-CN) was used for internal reference detection of samples. HRP-labeled IgG is recommended, such as A21020 HRP sheep anti-rabbit secondary antibody.
b. Diluted antibodies should be stored at 4℃ immediately after antibody incubation to facilitate subsequent reuse. At the same time of washing the HRP-labeled secondary antibody on the membrane, the luminous working liquid was prepared by mixing the two reagents at a ratio of 1:1 to prepare the working liquid.
c. If the imprinted film size is 1 cm2, 1-0.2 mL SuperKine™ Enhanced ECL Luminescence (Peak Grade) working fluid is recommended.
d. Incubate in Superkine ™ Enhanced ECL Working Fluid for 1-5 minutes.
e. Pinch the membrane with tweezers and gently contact the lower edge of the membrane with the filter paper to remove excess luminescent liquid from the membrane. Cover the imprinted film with transparent plastic packaging.
f.Use X-ray film exposure or a chemiluminescence imager to take pictures.

Experimental Results:

Superkine ™ Ultra Sensitive ECL Luminescence (Flyker Grade, BMU102-CN) Signal Stability and Durability Verification Data

Figure: Verification data for signal stability and persistence of SuperKine™ Ultra Sensitive ECL Luminescence Solution (Fyker grade, BMU102-CN).
L929 cell samples were diluted 50ug, primary antibody: mouse anti-NFKBP65 mAb, (ABM0017, 1:5000); Secondary antibody: Goat anti-mouse IgG, (A21010, 1:10,000); Exposure time 60S, PVDF membrane; Imaging system: Chemiluminescence Imager.

BMU103-EN SuperKine™ Enhanced Antibody Diluent

Application: Superkine ™ Enhanced Antibody Dilution Buffer is a versatile, ready to use Antibody Dilution solution for WB, IF, ICC, IHC, ELISA, and other immunoassay applications. The antibody diluent can be used for the dilution and preparation of various primary and secondary antibodies by adding specialized immune signal enhancement components, as well as specially developed immune blocking and stabilizing components, to enhance the signal strength of the immune assay, reduce the reaction background, and obtain the best immune assay results. This product contains no protein, phosphate, sodium azide or thiomersal preservatives and is suitable for most types of antibody dilution:

a. Pure chemical composition - no animal protein, no risk of antigen-antibody cross-reaction.
b. Signal enhancement - contains antibody-antigen binding enhancement and is phosphate free.
c. Quick time saving -- Strong results can be achieved within 1 hour of incubation, without overnight, realizing the dream of finishing WB in 1 day.
d. Good compatibility - compatible with horseradish peroxidase, alkaline phosphatase and biotin labeled secondary antibodies.
e. Safe and non-toxic - adding new preservatives, no sodium azide, non-toxic and healthy, safe to use.
f. In solution -- use without dilution.
g. Widely applicable -- suitable for a variety of immunoassay (WB, IHC, ELISA, IF).

Experimental results:

Superkine™ Enhanced Antibody Diluent (BMU103-CN) was compared to a generic antibody diluent (TBST) and a comparable competitor (Brand B, Brand A) in a WB test

Figure: Superkine™ Enhanced Antibody Diluent (BMU103-CN) was compared to a generic antibody diluent (TBST) and a comparable competitor (Brand B, Brand A) in a WB test.
Protein samples: HEK293 cells; Primary antibody (NFkB mouse monoclonal antibody, ABM0017,1:5000) was incubated at room temperature for 1 h and washed with TBST. The secondary antibody (horseradish peroxidase labeled goat anti-mouse IgG, A21020,1:10000) was incubated at room temperature for 1 h and washed with TBST. ECL luminescent solution (BMU102-CN) was incubated for 1-2 min, and then exposed for the same time for imaging.

References:
[1]. Inhibition of Sema4D/PlexinB1 signaling alleviates vascular dysfunction in diabetic retinopathy. EMBO Mol Med (2020)12:e10154
[2]. Distribution and Dynamic Changes in Matrix Metalloproteinase (MMP)-2, MMP-9, and Collagen in an In Stent Restenosis Process. European Journal of Vascular and Endovascular Surgery, Volume 61, Issue 4, April 2021, Pages 656
[3]. Circulating IGF-1 promotes prostate adenocarcinoma via FOXO3A/BIM signaling in a double-transgenic mouse model. Oncogene volume 38, pages6338–6353(2019)

 

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