Overview

  • Product nameAnti-PMP70 antibody
    See all PMP70 primary antibodies
  • Description
    Rabbit polyclonal to PMP70
  • Tested applicationsSuitable for: ICC/IF, IHC-P, ICC, 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)

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. PubMed: 17643102
IHC-P 1/500.
ICC Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

0.5ug/test



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

  • FunctionProbable transporter. The nucleotide-binding fold acts as an ATP-binding subunit with ATPase activity.
  • Sequence similaritiesBelongs 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 localizationPeroxisome membrane.
  • Information by UniProt
  • Database links
  • 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
    • Peroxisomal membrane protein 1 antibody
    • PMP 70 antibody
    • PMP70 antibody
    • PXMP 1 antibody
    • PXMP1 antibody
    • ZWS2 antibody
    see all

Anti-PMP70 antibody images

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

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

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

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

  • Flow cytometry analysis of PMP70 in 293T cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 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.

  • Flow cytometry analysis of PMP70 in HepG2 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 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.

  • Flow cytometry analysis of PMP70 in NIH-3T3 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 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.

  • 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?)
  • 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
  • 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.
  • 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.

References for Anti-PMP70 antibody (ab3421)

This product has been referenced in:
  • Sargent G  et al. PEX2 is the E3 ubiquitin ligase required for pexophagy during starvation. J Cell Biol 214:677-90 (2016). Read more (PubMed: 27597759) »
  • Haldar AK  et al. IRG and GBP host resistance factors target aberrant, "non-self" vacuoles characterized by the missing of "self" IRGM proteins. PLoS Pathog 9:e1003414 (2013). Read more (PubMed: 23785284) »

See all 12 Publications for this product

Product Wall

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.


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

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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Verified customer

Submitted Apr 15 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
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
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Abcam user community

Verified customer

Submitted Mar 08 2012

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...

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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 ...

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