Overview

  • Product name
  • Description
    Goat polyclonal to Met (c-Met)
  • Host species
    Goat
  • Tested applications
    Suitable for: ICC/IF, Neutralising, ELISA, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment (extracellular domain)(Human).

  • General notes

    The detection limit in immunoblotting for recombinant human HGF R is approximately 5 ng/lane under non-reducing and reducing conditions. Both the alpha and beta chains of HGF R are detected by this antibody under reducing conditions. The detection limit in ELISA for recombinant human HGF R is approximately 0.16 ng/well.

Properties

Applications

Our Abpromise guarantee covers the use of ab10728 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
ICC/IF Use at an assay dependent concentration.
Neutralising Use a concentration of 0.5 - 2 µg/ml. ab10728 has the ability to neutralize receptor-ligand interaction. Approximately 0.5-2 mg/mL of the antibody will block 50% of the binding of recombinant human HGF (5 ng/mL) to immobilized recombinant human HGF R/Fc Chimera (100 mL of a 1 mg/mL solution coated in each well) in an ELISA. 10 mg/mL of the antibody will block 90% of binding.
ELISA Use a concentration of 0.5 - 1 µg/ml.
WB Use a concentration of 0.1 - 0.2 µg/ml. Predicted molecular weight: 129 kDa.

Target

  • Function
    Receptor for hepatocyte growth factor and scatter factor. Has a tyrosine-protein kinase activity. Functions in cell proliferation, scattering, morphogenesis and survival.
  • Involvement in disease
    Note=Activation of MET after rearrangement with the TPR gene produces an oncogenic protein.
    Note=Defects in MET may be associated with gastric cancer.
    Defects in MET are a cause of hepatocellular carcinoma (HCC) [MIM:114550].
    Defects in MET are a cause of renal cell carcinoma papillary (RCCP) [MIM:605074]. It is a subtype of renal cell carcinoma tending to show a tubulo-papillary architecture formed by numerous, irregular, finger-like projections of connective tissue. Renal cell carcinoma is a heterogeneous group of sporadic or hereditary carcinoma derived from cells of the proximal renal tubular epithelium. It is subclassified into common renal cell carcinoma (clear cell, non-papillary carcinoma), papillary renal cell carcinoma, chromophobe renal cell carcinoma, collecting duct carcinoma with medullary carcinoma of the kidney, and unclassified renal cell carcinoma.
    Note=A common allele in the promoter region of the MET shows genetic association with susceptibility to autism in some families. Functional assays indicate a decrease in MET promoter activity and altered binding of specific transcription factor complexes.
    Note=MET activating mutations may be involved in the development of a highly malignant, metastatic syndrome known as cancer of unknown primary origin (CUP) or primary occult malignancy. Systemic neoplastic spread is generally a late event in cancer progression. However, in some instances, distant dissemination arises at a very early stage, so that metastases reach clinical relevance before primary lesions. Sometimes, the primary lesions cannot be identified in spite of the progresses in the diagnosis of malignancies.
  • Sequence similarities
    Belongs to the protein kinase superfamily. Tyr protein kinase family.
    Contains 3 IPT/TIG domains.
    Contains 1 protein kinase domain.
    Contains 1 Sema domain.
  • Domain
    The kinase domain is involved in SPSB1 binding.
  • Post-translational
    modifications
    Dephosphorylated by PTPRJ at Tyr-1349 and Tyr-1365.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • AUTS9 antibody
    • c met antibody
    • D249 antibody
    • Hepatocyte growth factor receptor antibody
    • HGF antibody
    • HGF receptor antibody
    • HGF/SF receptor antibody
    • HGFR antibody
    • MET antibody
    • Met proto oncogene tyrosine kinase antibody
    • MET proto oncogene, receptor tyrosine kinase antibody
    • Met proto-oncogene (hepatocyte growth factor receptor) antibody
    • Met proto-oncogene antibody
    • Met protooncogene antibody
    • MET_HUMAN antibody
    • Oncogene MET antibody
    • Par4 antibody
    • Proto-oncogene c-Met antibody
    • RCCP2 antibody
    • Scatter factor receptor antibody
    • SF receptor antibody
    • Tyrosine-protein kinase Met antibody
    see all

Images

  • ab10728 staining mouse muscle cells by ICC/IF.  Cells were PFA fixed and permeabilized in 0.5% Triton prior to blocking with 1% serum for 30 minutes at 25°C.  The primary antibody was diluted 1/50 and incubated with the sample for 1 hour at 25°C.  An Alexa Fluor® 488 conjugated chicken anti-goat antibody was used as the secondary.

    See Abreview

References

This product has been referenced in:
  • Morancho B  et al. Role of ADAM17 in the non-cell autonomous effects of oncogene-induced senescence. Breast Cancer Res 17:106 (2015). Read more (PubMed: 26260680) »
  • An N  et al. Activation of Pim Kinases Is Sufficient to Promote Resistance to MET Small-Molecule Inhibitors. Cancer Res 75:5318-28 (2015). Read more (PubMed: 26670562) »
See all 4 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Answer

Thank you for contacting us.


I contacted the lab and regrettably I can confirm, that this antibody has to our knowledge not been tested in vivo in nude mice or any other mice. For this reason we con not provide any related data. I am sorry that I cannot be of more help on this occasion.

I hope this information might be helpful to you nevertheless. Please do not hesitate to contact us if you need any more advice or information.

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Question

Dear Techserve, kindly receive customers reply below:

Thank You for Your reply! Please look below for the answers.

1. Has the protein transfer to the membrane and quality of the sample been assessed using a loading control?

We always use a loading control for western blot. The pdf document that I had sent to you earlier indicates beta actin as a loading control clearly.

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

Yes, it does. We are running several western blottings every day, so our secondary antibodies are almost daily tested by several colleagues.

The host species for the abcam c-MET primary antibody is goat. For beta actin primary antibody , the host species is goat as well. The secondary antibody used was the same for both these primary antibodies (donkey anti-goat). The beta actin bands are very clear, whereas there were no bands for c-MET. This clearly indicates that the current secondary antibody vial works.

Similarly, the rabbit secondary antibody used for flow cytometry has been in continuous use in our lab and the current vial works.

3. Increasing the antibody concentrations

Regarding the western blotting: if the signal is very mild or weak, then we can enhance the signal using a higher primary antibody concentration, but if we have no signal (even with the femto chemiluminiscence detection), then I personally have doubt that increasing the antibody concentration will help. With respect to flow cytometry, increasing the concentration of primary antibody, we have to increase the corresponding IgG control, which in turn may result in proportional higher signal. Nevertheless, I will take your suggestion and use a higher concentration of the antibodies.

In the meanwhile, could we kindly proceed with this decision of replacement of the antibodies.

Await your reply thankfully!

Read More
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry the products did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number ######.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Question
Answer

Thank you for taking the time to complete our questionnaire.

The details you have kindly provided will provide us with vital information for our monitoring of product quality.

I appreciate the time the customer have spent in the laboratory and understand their concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there very few tips to provide on this occasion to help improve the results from either of these antibodies. I can suggest it is possible you may have regrettably received a bad vial on this occasion.

However, I can recommend it would be important to consider the following:

ab10728 WB
1. Has the protein transfer to the membrane and quality of the sample been assessed using a loading control?
2. Is the current vial of secondary antibody working well with other primary antibodies?
3. I can suggest to try increasing the antibody concentration, for example 0.5 or even 1 ug/ml. The dilution provided on the datasheet is a guideline only. Increasing the concentration may help increase the signal.

ab2349
1. Antibodies are usually used at quite a high concentration in flow cytometry, often at a dilution between 1:10 to 1:50. Has this been tried? It may increase the signal obtained.
2. Is the current vial of secondary antibody working well with other primary antibodies?

Alternatively, or if these tips do not work, I am pleased to offer you a free of charge replacement or credit note for each antibody in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More

Answer

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

I would like to reassure you that our antibody are tested and covered by our 6 month guarantee for the tested applications and species listed on the individual datasheets. 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 appreciate your concerns and it is regrettable the results have not been successful. Before deciding how to proceed in this case, 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 have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.

WESTERN BLOT:


Order Details
Antibody code:

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number:



General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, wrong band size, more bands, no band etc.)


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


Amount of protein loaded


Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method (ECL, ECLPlus etc.)


Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the Western?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?


Additional Notes:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.




FLOW CYTOMETRY

Order Details
Antibody code:


Problem:
Choose: Non-specific staining No signal or weak signal High background


Lot number

Purchase order number
or preferably Abcam order number


General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, low signal, no signal etc.)


Sample (Species/Cell type/Cell line etc.)


Sample preparation for single cell preparation (Buffer etc.)


Number of cells used


Permeabilization, fixation step


Blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method


Positive and negative controls used (please specify)




Optimization attempts (problem solving)
How many times have you tried the FACS?



Have you run a "No Primary" control?


Yes No
Do you obtain the same results every time?
Yes No

What steps have you altered?


Additional Notes:


Data:
We would appreciate if you are able to provide any data/histograms which will help us to assess the results

Read More

Answer

Thank you for your reply. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. Unfortunately, the application you are planning (WB when the receptor c-Met is bound by HGF)is not eligible for this program.

I therefore would like to offer you two special discounts as a special exception: 5% off when you order ab10728 and free shipping.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Pleaselet me know if your are planning toorder this antibody and I will be happy to provide the discount codes.

Read More

Answer

Thank you for your inquiry.

This antibody can be used to neutralise the c-Met:HGF interaction - At 0.5-2 μg/mL the antibody will block 50% of the binding of 5 ng/mL recombinant human HGF to 0.1 μg immobilized recombinant human HGF R/c-MET Fc chimera. >90% of the binding will be blocked by 10 μg/mL of the antibody.

To the best of my knowledge this antibody has not been tested against HGF-bound receptor. As the antibody blocks the interaction it is possible that it binds at the same site as HGF and so might in turn be blocked by the presence of HGF. Conversely as this is a polyclonal antibody there may be sufficient non-HGF binding site epitope recognition to allow binding.

Unfortunately as this has not been tested I could not say for sure.

I am sorry I could not be of more help, but please do not hesitate to contact us if you require any further information or assistance.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (muscle cells)
Specification
muscle cells
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Nov 01 2007

Answer

Thank you for your enquiry. For ab10728, the immunogen was from the extracellular domain. It was not tested against a cell line. The neutralization assay is described as follows. Neutralization of Receptor-Ligand Interaction: Approximately 0.3-1.0 ug/ml of this antibody will block 50% of the binding of 5 ng/ml of rhHGF to immobilized rhHGF R/Fc chimera (100 uL of a 1 ug/ml solution was coated in each well) in a functional ELISA assay. For ab10681, it was not tested against a cell line. The neutralization assay is described as follows. Neutralization of Receptor-Ligand Interaction: Approximately 0.5-2.0 ug/ml of this antibody will block 50% of the binding of 5 ng/ml of rhHGF to immobilized rhHGF R/Fc chimera (100 uL of a 1 ug/ml solution was coated in each well) in a functional ELISA assay. If you have any additional questions, please contact us again.

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