Overview

  • Product name
  • Description
    Rabbit polyclonal to Flotillin 1
  • Host species
    Rabbit
  • Specificity
    This target has been reported to show both cytoplasmic and nuclear localisation (PMID: 15713644).
  • Tested applications
    Suitable for: IHC-P, ICC/IF, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human
    Predicted to work with: a wide range of other species
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Flotillin 1.

    Read Abcam's proprietary immunogen policy (Peptide available as ab41926.)

  • Positive control
    • ab41927 gave a positive result in Human Lung tissue lysate, Human Brain tissue lysate and Hela whole cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab41927 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 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/200. Fix cells with methanol
WB Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).Can be blocked with Human Flotillin 1 peptide (ab41926).
IP Use at an assay dependent concentration.

Target

Images

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Flotillin 1 knockout HAP1 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab41927 observed at 47 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab41927 was shown to specifically react with Flotillin 1 in wild-type HAP1 cells as signal was lost in Flotillin 1 knockout cells. Wild-type and Flotillin 1 knockout samples were subjected to SDS-PAGE. ab41927 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 µg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • ab41927 (1/200) staining Flotillin 1 in HeLa cells (green). Cells were fixed with Methanol and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.

    See Abreview

  • All lanes : Anti-Flotillin 1 antibody (ab41927) at 1 µg/ml

    Lane 1 : Lung (Human) Tissue Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : Human brain tissue lysate - total protein (ab29466)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 47 kDa
    Observed band size: 47 kDa

  • ab41927 (1/100) staining Flotilin 1 in paraffin embedded - Human (anal cancer), Mouse (brain) and Rat (spleen) tissues. Tissue sections underwent paraformaldehyde fixation, heat mediated antigen retrieval using Citrate Buffer pH 6.0 and were blocked using sequential peroxidase and protein block (prediluted). Please refer to abreview for further experimental details.

    See Abreview

  • Flotillin 1 immunoprecipitation from Human HeLa whole cell lysate using ab41927. 750 µg of cell lysate was incubated with primary antibody (1/500) and matrix (Protein A) and incubated for 16 hours at 4°C.

    Lane 1 : anti-Flotillin 2 IP (using non-Abcam antibody)
    Lane 2 : anti-Flotillin 2 IP (using ab41927)
    Lane 3 : Input (HeLa cell lysate)

    Western blot detection : mouse anti-Flotillin 1 antibody + HRP donkey anti-mouse polyclonal secondary antibody

    See Abreview

  • ICC/IF image of ab41927 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab41927 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • All lanes : Anti-Flotillin 1 antibody (ab41927) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Human brain tissue lysate - total protein (ab29466)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 47 kDa
    Observed band size: 50 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 35 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 4 minutes

References

This product has been referenced in:
  • Xie JC  et al. Hypoxia increases amyloid-ß level in exosomes by enhancing the interaction between CD147 and Hook1. Am J Transl Res 10:150-163 (2018). Read more (PubMed: 29423001) »
  • Real JM  et al. Exosomes from patients with septic shock convey miRNAs related to inflammation and cell cycle regulation: new signaling pathways in sepsis? Crit Care 22:68 (2018). Read more (PubMed: 29540208) »
See all 21 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Application
IHC - Wholemount
Sample
Xenopus laevis Embryo (st.11.5, dorsal view)
Specification
st.11.5, dorsal view

Abcam user community

Verified customer

Submitted Mar 26 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HeLa)
Total protein in input
750 µg
Specification
HeLa
Immuno-precipitation step
Protein A

Prof. Ritva Tikkanen

Verified customer

Submitted Jan 11 2013

Answer

I am very pleased to hear you would like to test both ab41927 and ab96507 in immunoprecipitation. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for immunoprecipitation and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.

For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways:

1) Call to place your order and mention the code to our customer service department;
2) Include the code in your fax order;
3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you for contacting us.

I'm sorry to hear that the antibody is not working anymore after the -80 degree storage.
As we discussed over the phone, both species are untested.

If you could test the antibody in human samples - a tested species - and then we can resolve the problem as we discussed: either with a free of charge replacement or a testing discount code.

I look forward to hear back from andyou assist you further.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More
Application
Immunocytochemistry
Sample
Mouse Cell (RAW 264.7)
Specification
RAW 264.7
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton-X100 in 2% BSA for 15min
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C

Dr. Mahesh Shivananjappa

Verified customer

Submitted May 24 2012

Question
Answer

DISCOUNT CODE: ***
Expiration date: May 23rd, 2012

I am very pleased to hear you would like to accept our offer and test ab41927 in macaque. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview formacaque and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you for your enquiry. flotillin1 is a peripheral membrane protein that is associated with caveolae. As such I believe it will be a cytoplasmic (internal) epitope that is detected by this antibody. However you may wish to consult the caveolae literature. I hope this is helpful. Please contact me again if you have any further questions.

Read More

Answer

Thank you very much for your interest in ab41927. To our knowledge, ab41927 has not been tested in Flow cytometry. Therefore, I can offer a discount off a future purchase if you buy ab41927 now, test it in Flow cytometry and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free PRIMARY ANTIBODY. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab41927 in Flow cytometry. I will then send a discount code. This code must be issued before purchasing ab41927 so please wait for my reply before ordering. 2. Purchase ab41927 either by phone, fax, or online (www.abcam.com). 3. Test it in Flow cytometry. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any PRIMARY ANTIBODY ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab41927 turns out to be unsuitable for Flow cytometry, you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Platelets)
Loading amount
25 µg
Specification
Platelets
Gel Running Conditions
Reduced Denaturing (4-15%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Dr. Mahesh Shivananjappa

Verified customer

Submitted Oct 31 2011

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Xenopus laevis Cell (embryos)
Specification
embryos
Fixative
Methanol
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 23 2011

1-10 of 15 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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