Overview

  • Product name
  • Description
    Rabbit polyclonal to FIGN
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    antigen sequence: IIVQLLSQHN YCLNDKEFAL LVQRTEGFSG LDVAHLCQEA VVGPLHAMPA TDLSAIMPSQ LRPVTYQDFE NAFCKIQPSI SQKELDMYVE W, corresponding to amino acids of Human FIGN.

  • Positive control
    • Human corpus unterine tissue.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    pH: 7.20
    Preservative: 0.02% Sodium azide
    Constituents: 59% PBS, 40% Glycerol
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab122238 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
IHC-P 1/1000 - 1/2500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 - 4 µg/ml.

Recommend PFA Fixation and Triton X-100 treatment

Target

Images

  • Immunofluorescent staining of Human cell line U-2 OS shows positivity in plasma membrane and cytoplasm. Recommended concentration of ab122238 1-4 µg/ml. Cells treated with PFA/Triton X-100.
  • ab122238, at 1/1000 dilution, staining FIGN in paraffin-embedded Human corpus uterine tissue by Immunohistochemistry.

References

ab122238 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Thank you for your response.

I would expect ab31850 to give clearer results in WB, even once the many isoforms are taken into consideration. I would be happy to offer you a free of charge replacement, credit, or refund for this antibody as well. Please let me know which antibodies you would like to receive as replacements and I will be happy to process your request.


For the FIGN antibody ab122238, I would recommend ensuring that you are performing heat mediated antigen retrieval with Citrate buffer, pH 6. In order to increase the signal, it could help to lengthen your antigen retrieval time. Can you please let me know what type of tissue sections you were testing?

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Question

Thank you for getting back to me. Sorry for the delay in getting this information to you. We have actually been testing a third antibody from you that is also giving us the same problem - anti–GPSM2 antibody (a b84571). Hopefully,
you will be able to assist us.
Below is the information you requested. I've included the information as well for the anti-GPSM2 antibody.

1) What was your original order or PO number?

Anti-spastin antibody (ab31850) - ***
Anti-FIGN antibody (ab122238) - ***
Anti-GPSM2 antibody (ab84571) - ***

2) Could you please provide a description of the problem (no signal, high background, incorrectstaining, etc) or an image of your results?
SPASTIN – many bands of incorrect size. None of the observed bands disappeared or decreased in intensity in cells transfected with siRNAs against Spastin.
FIGN – no signal
GPSM2 – no signal
The mRNAs for all these genes were detected by RT-PCR in the cells that were used for the WB.

3) What type of samples were you testing?
Which species were they from, what tissue type or cell line, and how were they prepared?
For example, what lysis buffer was used in WB, were the samples reduced and denatured, and how much protein was loaded?
For IF, how were the cells fixed and permeabilized?
Proteins extracts from RPE-1 (human), HeLa (human) and NIH3T3 (mouse) were used to test the FIGN and SPASTIN antibodies.
Proteins extracts from HeLa (human) were used to test the GPSM2 antibody.
The protein extracts were prepared with the following lysis buffer: 50mM HEPES, 100mM KCl, 0.3% NP-40, 10% glycerol, 1mM EGTA, 1mM MgCl2 + protease inhibitors and
DTT.
The protein extracts were quantified by the Bradford assay and 25mg were loaded/lane in 10% SDS-PAGE after
being incubated at 95ºC for 5 min.

4) What were the dilutions and incubation times for the block, primary, and secondary antibodies?
What blocking agent and secondary antibody were used?
The proteins were transferred to PVDF membranes. The membranes were blocked for 1h in 5% Skimmilk in TBS-Tween. The membranes were incubated with the primary antibodies in
blocking solution over night at 4ºC with constant agitation. For each antibody the following dilutions were tested: 1:100; 1:250; 1:500; 1.1000.
After the incubation with the primary antibodies the membranes were washed 3 times for 10 minutes with TBS-Tween. The membranes were then incubated with HRP- conjugated secondary
antibodies for 1h at room temperature. After this incubation the membranes were washed 3 times with TBS-Tween. Finally the membranes were incubated with ECL and exposed.

If you require any additional information please let me know. Any help you can provide would be great.

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Answer

Thank you for your reply.

For the Spastin antibody ab31850, could you please provide an image of your blot or let me know what size bands you are observing? Spastin has several known isoforms that could account for multiple bands between 54-67 kDa. Regarding the lack of change with transfected cells and siRNA, can you tell me if the transfected Spastin was full length and from human? What siRNA were you using, and was the knock-down validated by RT-PCR?

For the GPSM2 antibody, the protocol you provided seems fine and we would definitely expect you to get good signal using this method in HeLa cells by WB and ICC/IF. I would be happy to offer you a free of charge replacement, credit, or refund for this antibody.

The FIGN antibody ab122238 has not been validated for use in WB or ICC/IF on cultured cells. Have you tried this antibody in IHC-P? It is possible that it may work in WB or ICC/IF, but we do not have any information about how the antibody would work for those applications. To increase your signal, you could try preparing your samples with RIPA buffer, which is more effective at releasing nuclear proteins. You could also try increasing your ECL exposure time or your secondary antibody concentration.

I hope this helps, if not, please let me know and I will be happy to help you further.

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Answer

Thank you for contacting us. I am sorry that these antibodies are not working in WB and IF. We will be happy to refund or replace the antibodies if they are being used in a tested species and application, and have been purchased in the past six months. In order to help our QC process, could you please provide some additional information?

1) What was your original order or PO number?

2) Could you please provide a description of the problem (no signal, high background, incorrect staining, etc) or an image of your results?

3) What type of samples were you testing? Which species were they from, what tissue type or cell line, and how were they prepared? For example, what lysis buffer was used in WB, were the samples reduced and denatured, andhow much protein was loaded? For IF, how were the cells fixed and permeabilized?

4) What were the dilutions and incubation times for the block, primary, and secondary antibodies? What blocking agent and secondary antibody were used?

I will be happy to assist you further once I have this additional information.

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

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