Western blot test: 0.1-1 μg/ml
IHC, ICC, FACs and IP test: 1-5 μg/ml
Elisa test: 0.05-0.2 µg/ml
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Western blot test: 0.1-1 μg/ml
IHC, ICC, FACs and IP test: 1-5 μg/ml
Elisa test: 0.05-0.2 µg/ml
Please read and carefully follow the manual instructions on how to preserve each antibody product.
Typically, store antibodies at 2-8℃ for short term storage and store at -20℃ for long term storage. Aliquot and store before use to avoid repeated freeze-thaw cycles.
The following antibody products need to be stored in the dark.
To ensure antibody quality between different batches, our QA tests and compares sensitivity and specificity between batches
Polyclonal Antibodies
Large quantities of Polyclonal Antibody are relatively quick and inexpensive to produce compared to Monoclonal Antibodies. They are non-specific in that they are capable of recognizing multiple epitopes on any one antigen.
Advantages
Can help increase the WB signal as the antibody will bind to more than one epitope.
Due to recognition of multiple epitopes, polyclonal antibodies can give better results in IP/ChIP assays.
More tolerant of minor changes in the antigen, e.g., polymorphism, heterogeneity of glycosylation, or slight denaturation.
Useful when the nature of the antigen is unknown.
Inexpensive to produce and timeline is short.
Disadvantages
More prone to batch to batch variability.
Multiple epitopes make it important to check immunogen sequence for any potential cross-reactivity.
Monoclonal Antibodies
Advantages
Compared to polyclonal antibodies, homogeneity of monoclonal antibodies is very high.
If experimental conditions are kept constant, results from monoclonal antibodies will be highly reproducible between experiments.
All batches will be identical and specific to just one epitope which is a particular advantage when manufacturing procedures must be standardized e.g. clinical tests and therapeutic treatments.
The high specificity of monoclonal antibodies decreases background noise and cross-reactivity, helps provide reproducible results and ensure efficiency in affinity purification.
Specific antibody characteristics can be identified and selected e.g. sensitivity requirements and cross reactivity levels can be specified and screened to identify cell lines exhibiting the required characteristics.
Hybridomas provide an immortal cell line with the ability to produce unlimited quantities of highly specific antibodies.
Disadvantages
Antibodies may be too specific (e.g. less likely to detect across a range of species).
More vulnerable to the loss of epitope through chemical treatment of the antigen than polyclonal antibodies. This can be offset by pooling two or more monoclonal antibodies to the same antigen.
Expensive to produce and timeline is long for hybridomas.
The brightness of fluorophore and the antibody's sensitivity are two factors that influence detection sensitivity. THE™ Elite antibody series have the best sensitivity on the market. The brighter the fluorophore is, the better the overall detection. GenScript's iFluor antibody series are newly developed fluorescent compounds conjugated to THE™ Elite antibody series. Compared with traditional fluorescent dyes, the iFluor series are brighter and have better stability.
GenScript has four types of fluorophore conjugated His-tag antibodies for FACS analysis, including A01620 (FITC), A01800 (iFluor488), A01801 (iFluor555) and A01802 (iFluor 647). The iFluor series are recommended for FACS analysis because they have a higher sensitivity than FITC conjugated antibody. The following is the wavelength range of GenScript's iFluor antibodies
Cat. No. |
Conjugate |
Ex (nm) |
Em (nm) |
|---|---|---|---|
A01800 |
iFluor 488 |
491 |
514 |
A01801 |
iFluor 555 |
559 |
569 |
A01802 |
iFluor 647 |
654 |
674 |
According to our FACS analysis results, the sensitivity of A01802 (THE™ His Tag Antibody [iFluor 647], mAb, Mouse) is slightly higher than the A01800 (THE™ His Tag Antibody [iFluor 488], mAb, Mouse). If the His Tag number is low, A01802 is recommended.
For A00186, a starting point of 1 µg/ ml is recommended.
The catalog No. is A00186 for the anti-His mAb in the L00436 ELISA Kit. The recommended concentration for WB is 0.1-0.2 µg/ml.
Catalog No. A00174 is a rabbit anti-His tag antibody. It has been tested for internal His-tag recognition, but not for Flow Cytometry.
For Flow Cytometry, the following antibodies are recommended. All of them can recognize His-tag fused on the internal site of a protein.
Can A01795 (THE™ PEG Antibody, mAb, Mouse) recognize PEG5
We have not yet tested PEG5. We tested PEG12 with positive results. One of our clients tested PEG2 and observed good results.
Can I use A01795 (THE™ PEG Antibody, mAb, Mouse) in building an affinity column.
In most cases it should be OK to conjugate an antibody (IgG/IgM/IgY) to agarose resin. Our PEG antibody could be used for this conjugation.
Do you have any information on antibody stability after reconstitution? I'm concerned that an IgM antibody may have fragmented during the lyophilization process or became aggregated upon reconstitution.
As mentioned in the datasheet, THE™ PEG Antibody, mAb, mouse should be stored in its lyophilized form until use. It remains stable in lyophilized form if stored at -20°C or below. The reconstituted antibody can be stored for 2-3 weeks at 2-8°C or for up to 12 months at -20°C or below. Avoid repeated freeze-thaw cycles.
Reconstitute the lyophilized product with deionized water (or equivalent) to make an antibody concentration of 0.5 mg/ml. For long-term storage, it is recommended to add glycerol to a final concentration of 50% (v/v) in the reconstituted antibody solution.
When using [HRP] tagged primary antibodies there are some variables that may affect its binding with a secondary antibody.
Catalog No. A00166 (Goat Anti-Human IgG Antibody (H&L) [HRP], pAb) recognizes human IgG, but it has not yet been tested for cross-reactivity with other species.
Catalog. No A00160 (Goat Anti-Mouse IgG Antibody (H&L) [HRP], pAb) has not yet been tested for cross-reactivity with rabbit.
The immunogen of A01622 is human GAPDH protein from erythrocytes. A01622 detects endogenous levels of human, pig, cow, goat, and fish GAPDH. This product has not been tested for other species.
A00191 (GAPDH Antibody, pAb, Goat) specifically reacts with GAPDH from cell and tissue lysates of human, mouse, rat, goat, rabbit, chicken, hamster, bovine, swine, fish, and E. coli.
The MW of the target protein is about 18 kDa, so why is the band in the WB test picture almost 60 kDa.
The target protein tested is 57 KDa because it is the recombinant protein, not the naturally occurring protein with the molecular weight of 18 KDa.
What is the fusion partner
From the COA file, you can see that the fusion partners for this protein are calbindin, Trx tag and Strip-Ⅱtag.
Is there any cross-reactivity between the fusion partner and PIN1
According to multiple sequence alignment results, there is only one sequence in the recombinant protein with the peptide antigen for immunization. Based on these results there should be no cross-reactivity between the fusion partner and PIN1.
How is PIN1 fused to its partner
PIN1 is fused to its partner at the N terminal.
