Overview

  • Product name
    Anti-PLGF antibody - N-terminal
    See all PLGF primary antibodies
  • Description
    Rabbit polyclonal to PLGF - N-terminal
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ELISA, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human PLGF (N terminal). Highly purified N-terminal 20 aa synthetic peptide (Human).
    Database link: P49763

  • Positive control
    • WB: Purified, recombinant PLGF-1/-2. ICC/IF: HepG2 cells.
  • General notes

    According to Swissprot P49763; PLGF exists in three isoforms PLGF 1, 2 and 3. The N-terminal immunogen sequence of ab9542 is present in all isoforms so extra bands might be observed near the band of interest.

Properties

Applications

Our Abpromise guarantee covers the use of ab9542 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ELISA Use a concentration of 1 - 5 µg/ml.

Allows the detection of 1.0 - 2.5 ng/well of recombinant human PLGF-1 and PLGF-2.

WB Use a concentration of 1 - 4 µg/ml.

It will detect 25 - 50 ng/lane of recombinant human PLGF-1 (17-18 kDa) and PLGF-2 (22-23 kDa) under reducing conditions. Dimers are detected at higher protein amounts/lane.

IHC-P Use at an assay dependent concentration. PubMed: 19336420
ICC/IF Use a concentration of 1 µg/ml.

Target

  • Function
    Growth factor active in angiogenesis and endothelial cell growth, stimulating their proliferation and migration. It binds to the receptor FLT1/VEGFR-1. Isoform PlGF-2 binds NRP1/neuropilin-1 and NRP2/neuropilin-2 in a heparin-dependent manner.
  • Tissue specificity
    While the three isoforms are present in most placental tissues, PlGF-2 is specific to early (8 week) placenta and only PlGF-1 is found in the colon and mammary carcinomas.
  • Sequence similarities
    Belongs to the PDGF/VEGF growth factor family.
  • Domain
    Isoform PlGF-2 contains a basic insert which acts as a cell retention signal.
  • Post-translational
    modifications
    N-glycosylated.
  • Cellular localization
    Secreted. The three isoforms are secreted but PlGF-2 appears to remain cell attached unless released by heparin.
  • Information by UniProt
  • Database links
  • Alternative names
    • D12S1900 antibody
    • Pgf antibody
    • PGFL antibody
    • PIGF antibody
    • Placenta growth factor antibody
    • Placental growth factor antibody
    • Placental growth factor, vascular endothelial growth factor related protein antibody
    • PlGF 2 antibody
    • PlGF antibody
    • PLGF_HUMAN antibody
    • PlGF2 antibody
    • SHGC 10760 antibody
    see all

Images

  • Western blot of chromatography purified, recombinant PLGF-1/-2 and VEGF165 immobilized to PVDF-membrane.
    PlGF antibody shows no cross-reactivity to VEGF165.

  • ICC/IF image of ab9542 stained HepG2 (human liver hepatocellular carcinoma cell line) cells. The cells were fixed in 4% formaldehyde for 10 minutes and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9542, 1 µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.

References

This product has been referenced in:
  • Chen X  et al. Differential expression of vascular endothelial growth factor angiogenic factors in different endometrial compartments in women who have an elevated progesterone level before oocyte retrieval, during in vitro fertilization-embryo transfer treatment. Fertil Steril 104:1030-1036 (2015). Read more (PubMed: 26143364) »
  • Beiswenger TR  et al. The effect of cigarette smoke extract on trophoblast cell viability and migration: the role of adrenomedullin. Reprod Sci 19:526-33 (2012). WB ; Human . Read more (PubMed: 22267538) »
See all 13 Publications for this product

Customer reviews and Q&As

1-10 of 22 Abreviews or Q&A

Answer

Your credit note ID is XXXXX. Please pass this number to your finance department and ask them to quote this number when ordering new products. Once order received we will automatically deduct the money from invoice.

I hope this information will be helpful. Should you have any question please do not hesitate to ask.

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Answer

Thank you for contacting us.

Your credit note ID is 23009.

I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

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Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from these PLGF antibodies.

In answer to your question, I would like to reassure you that as far as I can see from our records, the Epitomics antibody is not the same as the two from Abcam you mention. We monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of these antibodies.

I would like to reassure you that these antibodies are tested and covered by our 6 month guarantee for the tested applications and species listed on the datasheet. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I would like to send you a technical questionnaire. This will help you put the information we require together very easily. I would appreciate if you could confirm which application you have been using, so I can send the correct questionnaire.

I would appreciate if you are able to provide any image which would help us to assess the results.

Thank you for your time and cooperation. I hope we can help you and I look forward to hearing from you with the requested details.

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Answer

Thank you for your reply.

That would not be a problem. However, could you please provide me with the order number on which ab9542 was received?

Many thanks in advance for your cooperation in this case.

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Answer

Thank you for contacting us. xxxx is currently away form the office but I will try to help as best I can.

I am sorry to hear that the protocol tips provided have not resulted in an improvement of the Western blotting performed. I the customer would like, I can offer to arrange for a replacement with an antibody such as ab83906 or ab97618. Alternatively, I can arrange for a credit note to be issued.

In order to arrange for either of these outcomes, could you please confirm the order numberon which the antibody ab9542 was ordered.

I look forward to hearing how the customerwould like to proceed.

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Answer

Thank you for providing this further information.

I appreciate your cooperation and understand your concerns. It is regrettable the results have not been successful. In order to help with our investigation of this case before deciding how to proceed, I would appreciate if you are able to provide more detail as requested in the previous email. This information will also be helpful to our quality monitoring.

2. Could you confirm if the quality of the sample and transfer to the membrane been assessed using a loading control?

A loading control is an antibody to a housekeeping protein such as GAPDH or actin.A loading control is commonly used in most western blots todetermine if the transfer to the membrane has been successful, that the loading between samples is even and that the sample is of good quality.

http://protocolsonline.com/protein-science/electrophoresis/loading-controls/

https://www.abcam.com/index.html?pageconfig=resource&rid=275

https://www.abcam.com/index.html?pageconfig=resource&rid=265

I would appreciate if you could confirm if a loading control has been included and what the results were?

Many thanks for yourcontinued cooperation.I will be pleased to provide a free of charge replacement or credit note if the antibody is not working with the suggestions provided.

Thank you for your time. I look forward to hearing from you with the requested information and hope we can resolve this case as soon as possible.

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Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

I would like to reassure you that ab9542 is tested and covered by our 6 month guarantee for use inWB and in human samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would like to offer some suggestions to help optimize the results. I would also appreciate if you can confirm some further details:

1. Could you provide details of which lysis buffer was used? I can recommend to use RIPA if this has not already been tried. This should provide a suitable protein preparation. Then, add sample buffer containing SDS and mercaptoethanol and heat to 95oC for 5 minutes to reduce and denature.

2. Could you confirm if the quality of the sample and transfer to the membrane been assessed using a loading control?

3. Was the overnight incubation at 4oC? This can often provide more specific and efficient staining.

4. To increase the signal, I suggest to try increasing the antibody concentration to 4 ug/ml, as recommended on the datasheet.

5. Themembrane provided in the image looks 'spotty'. I canrecommend to ensure the antibodyis mixed well with a pipette and to consider filtering theblocking agent.

6. Is the current vial of secondary antibody working well with other primary antibodies?

7. As far as I am aware from a literature search, the HUVEC cells express a receptor for PLGF protein, but I am not able to find anything to confirm the expression level of PLGF protein itself from these cells. Do you have any information to confirm the expression level of PLGF in HUVEC?

8. In this case, I can suggest it would be beneficial to include a positive control sample such as human placenta and brain tumor tissue.

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

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Answer

You email has been forward to me by my colleague.

The IHC-P dilution, 1/1000 is for guidelines purpose only; you can use this dilution as a start however if you observe faint staining then you might need more concentrated antibody e.g. 1/500 dilution and if high background or intense staining is observed then you might need more diluted antibody e.g 1/2000. The final otimal conditions has to be determined by the end user which changes with tissue sections and protocol etc.

I hope this information will be helpful. Should you have any other question please do not hesitate to ask.

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Answer

Thank you for your email.

- No primary control, is a negative control which is used to check the non specific binding of secondry antibody.

Please check my emails in comparison to the literature. http://www.uniprot.org/uniprot/P49763

The PLGF which exist as 3 isoforms is found as both homodimer and monomer. The size of monomer is 25 kDa and dimer will be ˜50 kDa. However because the protein is heavily glycosylated the observed band size will be slightly higher than the predicted band size. The antibody suppose to detect both the monomeric as well as dimeric form; the observance of dimer and monmer lies on the reducing conditions, if the lysates are well reduced with equal amount of LDS buffer there should be a one band and if the LDS buffer is not enough and heating si not enough for denaturing then the image will be dirty and there will be more bands.

The band at 50kDa could be the dimer or it could be the non specific band, to prove the results experiments always needs repetition. I am sorry we can not make any clear cut judgement based on only single image provided. You may have to repeat the experiment however if second time the same banding pattern is observed then we can antibody could be a faulty one.

This is very simple research procedure, I don't think it is confusing in anyway.

I am sorry this is all we have to say about the additional bands and we have no further recommendations to make. If you are not completely satisfied by the antibody please let me know I will refund the money you paid for.

I hope this information is never the less helpful. Should you have any question please do not hesitate to contact me.

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Answer

Thank you for your email.

The only reason for double band is dimer of PLGF protein. Please re do the WB with following suggestions;

- Heat lysates with sample buffer 10 minutes at 100C.
- Load 30ug protein per well
- Try no primary control
- Please make sure the lysates are fresh]

We unfortunately do no have nay other suggestions to make so the antibody should work with this protocol. In case the results stays the same please contact me for further assistance.

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1-10 of 22 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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