PLXND1 Polyclonal Antibody (E-AB-13524)

Q1:What is the concentration of primary antibody?

Our antibodies are concentrated, and the final concentration of the antibodies is not only related to the concentration of the antibodies themselves, but also to the abundance of the target protein in the sample. Therefore, the recommended dilution ratio for the antibodies provided in our instructions is a range, and the final dilution ratio needs to be determined through pre-experiments.You can also consult our technical team to get a recommendation of a suitable dilution ratio for your experimental conditions.

Q2:What does IHC mean on the datasheet? Is it a frozen tissue section? How is it different from IHC-F?

IHC is an immunohistochemical experiment, and the sections used generally include IHC-P (paraffin tissue section) and IHC-F (frozen tissue section).

Q3:The molecular weight in the literature is different from that in the manual. Why are there several molecular weights for the same index?

Because the same protein in different types of cells may have different post-transcriptional splicing bodies and post-translational modifications, such as glycosylation, ubiquitination, etc., the molecular weight is not the only constant. In addition, the protein You can find on the official Uniprot website that the molecular weights of different subtypes of proteins are different. Therefore, there will be a certain difference between the molecular weight found in the literature and the molecular weight of the antibody in the instructions. (You can check some literature references to confirm the molecular weight of the target protein in the target sample).

Q4:In IHC experiment, what are the conditions for antigen retrieval? Should I use EDTA or potassium citrate?

The repair conditions are generally high temperature and high pressure or microwave heating repair. For example: Antigen retrieval (high pressure method): Add 10 mmol/ml citrate buffer (pH 6.0) sufficient to submerge the slices in the pressure cooker (preparation: dissolve the antigen retrieval solution in the corresponding volume of ddH2O, mix well), and heat When it boils, place the slices on a heat-resistant plastic slicing rack, put them into the pot, cover the lid, fasten the pressure valve, continue heating, set the holding pressure for 4 minutes, open the vent valve to deflate after the time is up, and the pressure returns to zero. Open the lid, take out the inner pot and let it cool to room temperature. After the solution has cooled to room temperature, take out the slices (about 30 minutes). Antigen retrieval (microwave method): Add 10 mmol/ml citrate buffer (pH 6.0) sufficient to submerge the slices in a beaker (preparation: dissolve the antigen retrieval solution in the corresponding volume of ddH2O, mix well), and heat to boiling , place the slices on a heat-resistant plastic slice rack, put them into a beaker, heat over medium-low heat for 30 minutes, take them out, leave them to cool at room temperature, and take out the slices after the solution cools to room temperature (about 30 minutes). For the selection of IHC repair solution, it is recommended to refer to the antibody instruction manual, or refer to the following scheme: for human samples, it is preferred to use EDTA repair solution with pH 9.0 (clinical samples); for rat/mouse samples, it is preferred to use citric acid with pH 6.0 Repair fluid (scientific research sample).

Q5:If the activity of the antibody does not include the species of my experimental sample, will after-sales service be provided?

The species mentioned in the antibody instructions can be confirmed, but there is no guarantee that the antibody can be used in unverified species, even if the sequence homology is high. Whether an antibody will recognize proteins in samples from other species involves many factors and we cannot predict. If there is no other choice, you must consider purchasing antibodies for use in unverified species. We recommend that you compare the immunogen sequence with your target protein sequence. The higher the homology, the greater the possibility of success. If the antibody you purchased is for use in unverified species or experimental applications, product replacement or refunds are generally not available.

Q6:How to choose positive control for antibodies?

The use of positive control is the basis for determining whether the antibody and detection system work normally in an experiment, and it is also a basis for determining whether the protein to be detected exists in the sample to be detected! Especially when the detected sample is uncertain whether the protein to be detected is expressed. Therefore, when there is no positive result in the experiment, it is best to choose the appropriate positive control, and most of the instructions have recommended positive controls; Or you can search according to the SwissProt or Omnigene database in the instructions, these databases often list the tissues in which the protein is highly expressed, and these samples can be used as suitable positive controls.

Q7:How many films can I make with 20μl of antibody with IHC experimental application?

First of all, customers need to estimate the dosage of antibody according to the antibody instructions & the results of pre-experiments. Different samples will have different antibody dilution ratios due to different antigen expression. For example, in the IHC test of an antibody, the pre-test confirmation is calculated according to the dilution ratio of 1: 50. Under normal circumstances, one slice needs to add 100μl of diluted antibody, that is, one slice needs 2μl of antibody, and 20μl can make about 10 slices.

Q8:Can the antibody be used without the reactivity and application stated in the instructions?

General antibody instructions will list the types of experiments for which the antibody has been verified to be suitable (such as: WB, IHC, IP, IF, etc.). If the antibody instructions do not mention the application type, it does not mean that the antibody is not suitable. For this type of analysis application, it may be that we have not done the corresponding verification, or the verification sample is not very effective. Generally, if there is no application or the reactivity is not recommended for customers, please try to follow the verified ones listed in the instructions. Type to choose the right antibody for your experiment. If customers want to try it, they can consult technical support in advance or purchase small specifications for corresponding experiments. However, if the expected results are not achieved, product replacement or refunds are generally not available.