Anti-PMP70 antibody (ab3421)

Reviews (2)Q&A (4)References (29)

Overview

  • Product name

    Anti-PMP70 antibody
    See all PMP70 primary antibodies

  • Description

    Rabbit polyclonal to PMP70

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, IHC-P, Flow Cyt, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Hamster, Human, Non human primates

  • Immunogen

    Synthetic peptide corresponding to Rat PMP70 aa 644-659.
    Sequence:

    NYEFKKITEDTVEFGS


    (Peptide available as ab4965)

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

Properties

Applications

Our Abpromise guarantee covers the use of ab3421 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 1/600.
IHC-P 1/500.
Flow Cyt Use at an assay dependent concentration.

0.5ug/test

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
WB 1/500. Detects a band of approximately 70 kDa.Can be blocked with PMP70 peptide (ab4965). 70kDa band represents PMP 70 from peroxisome-enriched fractions from L cells.

Target

  • Function

    Probable transporter. The nucleotide-binding fold acts as an ATP-binding subunit with ATPase activity.

  • Sequence similarities

    Belongs to the ABC transporter superfamily. ABCD family. Peroxisomal fatty acyl CoA transporter (TC 3.A.1.203) subfamily.
    Contains 1 ABC transmembrane type-1 domain.
    Contains 1 ABC transporter domain.

  • Cellular localization

    Peroxisome membrane.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • 70 kDa peroxisomal membrane protein antibody
    • ABC 43 antibody
    • ABC D3 antibody
    • ABC43 antibody
    • ABCD 3 antibody
    • Abcd3 antibody
    • ABCD3 protein antibody
    • ABCD3_HUMAN antibody
    • ATP binding cassette sub family D (ALD) member 3 antibody
    • ATP binding cassette sub family D member 3 antibody
    • ATP-binding cassette sub-family D member 3 antibody
    • CBAS5 antibody
    • dJ824O18.1 (ATP-binding cassette, sub family D (ALD), member 3 (PMP70, PXMP1)) antibody
    • Peroxisomal membrane protein 1 (70kD) antibody
    • Peroxisomal membrane protein 1 (70kD, Zellweger syndrome) antibody
    • Peroxisomal membrane protein 1 antibody
    • PMP 70 antibody
    • PMP70 antibody
    • PXMP 1 antibody
    • PXMP1 antibody
    • ZWS2 antibody
    see all

Images

  • Western blot - Anti-PMP70 antibody (ab3421)
    Western blot - Anti-PMP70 antibody (ab3421)
    All lanes : Anti-PMP70 antibody (ab3421) at 1/1000 dilution

    Lane 1 : Mouse lung cell lysate
    Lane 2 : Mouse liver cell lysate
    Lane 3 : HepG2 cell lysate

    Lysates/proteins at 25 µg per lane.

    Observed band size: 70 kDa
    why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMP70 antibody (ab3421)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMP70 antibody (ab3421)
    ab3421 (2µg/ml) staining PMP70 in human duodenum using an automated system (DAKO Autostainer Plus). Using this protocol there is apical cytoplasmic staining and staining of the endoplasmic reticulum in the epithelium.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab3421)
    Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab3421)

    ICC/IF image of ab3421 stained human HeLa cells. The cells were 4% PFA 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 (ab3421, 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 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). This antibody also gave a positive IF result in Hek293, HepG2 and MCF7 cells.

  • Flow Cytometry - Anti-PMP70 antibody (ab3421)
    Flow Cytometry - Anti-PMP70 antibody (ab3421)

    Flow cytometry analysis of PMP70 in 293T cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x106 cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab3421 at a dilution of 0.5 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

  • Western blot - Anti-PMP70 antibody (ab3421)
    Western blot - Anti-PMP70 antibody (ab3421)
    All lanes : Anti-PMP70 antibody (ab3421) at 1/500 dilution

    Lane 1 : Rat kidney whole cell extracts at 20 µg
    Lane 2 : Rat liver whole cell extracts at 20 µg
    Lane 3 : Mouse lung whole cell extracts at 20 µg
    Lane 4 : A431 whole cell extracts at 10 µg
    Lane 5 : U2OS whole cell extracts at 10 µg
  • Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab3421)
    Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab3421)

    Immunocytochemistry/Immunofluorescence analysis of PMP70 using ab3421 shows staining in HMVEC Cells.

  • Flow Cytometry - Anti-PMP70 antibody (ab3421)
    Flow Cytometry - Anti-PMP70 antibody (ab3421)

    Flow cytometry analysis of PMP70 in HepG2 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x106 cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab3421 at a dilution of 0.5 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

  • Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab3421)
    Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab3421)

    Immunocytochemistry/Immunofluorescence analysis of PMP70 using ab3421 shows staining in NS-1 Cells.

  • Flow Cytometry - Anti-PMP70 antibody (ab3421)
    Flow Cytometry - Anti-PMP70 antibody (ab3421)

    Flow cytometry analysis of PMP70 in NIH-3T3 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x106 cells/ml, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab3421 at a dilution of 0.5 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

  • Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab3421)
    Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab3421)

    Immunocytochemistry/Immunofluorescence analysis of PMP70 using ab3421 shows staining in p19 Cells.

  • Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab3421)
    Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab3421)

    Immunocytochemistry/Immunofluorescence analysis of PMP70 using ab3421 shows staining in A549 Cells.

References

This product has been referenced in:

  • Abed M  et al. The Gag protein PEG10 binds to RNA and regulates trophoblast stem cell lineage specification. PLoS One 14:e0214110 (2019). Read more (PubMed: 30951545) »
  • Morito K  et al. Gut microbial metabolites of linoleic acid are metabolized by accelerated peroxisomal ß-oxidation in mammalian cells. Biochim Biophys Acta Mol Cell Biol Lipids 1864:1619-1628 (2019). Read more (PubMed: 31351225) »
See all 29 Publications for this product

Publishing research using ab3421? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-6 of 6 Abreviews or Q&A

Question

it is noted in the figure legend that Dylight 488-conjugated goat anti-mouse IgG were used. I am not sure it is correct secondary antibodies since the primary antibodies are rabbit.

How were cells' permeabilized for flow?



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Answer

I'm sorry for the confusion here. The secondary should be referred to as an 'anti-rabbit IgG'.

Cells were permeabilized for flow by incubation with ice cold 90% methanol for 30 min on ice.

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Question

It seems this protein is intracellular but the flow data does not indicate use of a reagent to permeabilize cells. So I just want to clarify the sub cellular localization and the protocol. thanks.

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Answer


What should be noted in the figure legends is that these cells were permeabilized with 90% ice cold methanol for 30 minutes.

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Western blot abreview for Anti-PMP70 antibody

 Excellent
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (12 %)
Sample
Human Cell lysate - whole cell (neuroblastoma)
Specification
neuroblastoma
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 16°C
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Abcam user community

Verified customer

Submitted Apr 15 2014

Western blot abreview for Anti-PMP70 antibody - Peroxisomal Membrane Marker

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (HEK293)
Loading amount
100 µg
Specification
HEK293
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Mar 08 2012

Question

I would like to test this antibody in IP

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Answer

I am very pleased to hear you would like to accept our offer and test ab3421 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. 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.

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Question

Hi dears;   I want to do Immunopercipitation for PMP-70 in peroxisomes (mouse brain tissue). I have anti PMP-70, rabbit polyclonal (ab3421),abcam. Can I use it for IP or not? I did not see any application for IP in your products. please help me in this issue.   Best Regards

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Answer

Thank you very much for your interest in ab3421. To our knowledge, ab3421 has not been tested in IP. Therefore, I can offer a discount off a future purchase if you buy ab3421 now, test it in  IP 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 ab3421 in  IP. I will then send a discount code. This code must be issued before purchasing ab3421 so please wait for my reply before ordering. 2. Purchase ab3421 either by phone, fax, or online (www.abcam.com). 3. Test it in  IP. 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 ab3421 turns out to be unsuitable for  IP, 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.  

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