Overview

  • Product name
  • Description
    Rabbit polyclonal to PPAR delta
  • Host species
    Rabbit
  • Specificity
    This affinity purified antibody is directed against mouse PPAR delta protein.
  • Tested applications
    Suitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide:

    MEQPQEETPEAREE

    , corresponding to N terminal amino acids 1-14 of Mouse PPAR delta .

  • Positive control
    • 3T3 cell lysate HepG2
  • General notes

    OMIM link - 600409

Properties

Applications

Our Abpromise guarantee covers the use of ab8937 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 a concentration of 1 µg/ml.
WB 1/1000 - 1/4000. Detects a band of approximately 54 kDa (predicted molecular weight: 50 kDa).

Target

  • Function
    Ligand-activated transcription factor. Receptor that binds peroxisome proliferators such as hypolipidemic drugs and fatty acids. Has a preference for poly-unsaturated fatty acids, such as gamma-linoleic acid and eicosapentanoic acid. Once activated by a ligand, the receptor binds to promoter elements of target genes. Regulates the peroxisomal beta-oxidation pathway of fatty acids. Functions as transcription activator for the acyl-CoA oxidase gene. Decreases expression of NPC1L1 once activated by a ligand.
  • Tissue specificity
    Ubiquitous with maximal levels in placenta and skeletal muscle.
  • Sequence similarities
    Belongs to the nuclear hormone receptor family. NR1 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • FAAR antibody
    • MGC3931 antibody
    • NR1C2 antibody
    • NUC1 antibody
    • NUCI antibody
    • NUCII antibody
    • Nuclear hormone receptor 1 antibody
    • Nuclear receptor subfamily 1 group C member 2 antibody
    • Peroxisome proliferative activated receptor delta antibody
    • Peroxisome proliferator-activated receptor beta (PPAR-beta) antibody
    • Peroxisome proliferator-activated receptor beta antibody
    • Peroxisome proliferator-activated receptor delta antibody
    • PPAR beta antibody
    • PPAR-beta antibody
    • PPAR-delta antibody
    • PPAR-ß antibody
    • PPARB antibody
    • ppard antibody
    • PPARD_HUMAN antibody
    see all

Images

  • ICC/IF image of ab8937 stained HepG2 cells (ab7900). The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum (ab7481) / 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 (ab8937, 1µg/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) at a concentration of 1.43µM.

  • Rabbit polyclonal to PPAR delta (ab8937).
    Each lane contains 20 ug 3T3 cell lysate

    Lane 1: ab8937 1/500
    Lane 2: ab8937 1/1000

    Secondary antibody: Goat anti-Rabbit (HRP) (ab6721) at 1/2000

    Rabbit polyclonal to PPAR delta (ab8937). Each lane contains 20 ug 3T3 cell lysate (ab7179) Lane 1: ab8937 1/500 Lane 2: ab8937 1/1000 Secondary antibody: Goat anti-Rabbit (HRP) (ab6721) at 1/2000

References

This product has been referenced in:
  • Mandai S  et al. WNK1 regulates skeletal muscle cell hypertrophy by modulating the nuclear localization and transcriptional activity of FOXO4. Sci Rep 8:9101 (2018). Read more (PubMed: 29904119) »
  • Mukherjee S  et al. Boosting efferocytosis in alveolar space using BCG vaccine to protect host against influenza pneumonia. PLoS One 12:e0180143 (2017). WB ; Mouse . Read more (PubMed: 28686604) »
See all 10 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (Colorectal cancer HCT116)
Loading amount
10 µg
Specification
Colorectal cancer HCT116
Gel Running Conditions
Non-reduced Denaturing (10% Gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Ms. Injoo Hwang

Verified customer

Submitted Nov 09 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - nuclear (Liver)
Loading amount
20 µg
Specification
Liver
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Jul 10 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Brain)
Loading amount
30 µg
Specification
Brain
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 07 2008

Question

AbCam Technical Support: I have written the answers below in CAPITAL letters beneath each issue. Thanks for any help you can provide > Also, please include the lot number (located on your vial) 68633 and the Abcam order number or purchase order number that was used. 1. Please describe the problem (high background, wrong band size, more bands, no band etc). THE PROBLEM IS THAT THE BAND THAT STAINS MOST INTENSELY IS LOCATED AROUND 70-75 kDA. MULITPLE OTHER BANDS APPEAR WITH LONGER EXPOSURE TIMES AND IT IS UNCLEAR WHICH BAND, IF ANY, REPRESENTS PPAR b. 2. On what material are you testing the antibody in WB? Species? HUMAN Cell extract/ Nuclear extract? CYTOPLASMIC AND NUCLEAR PROTEIN EXTRACTS FROM TWO DIFFERENT HUMAN CELL LINES Purified protein? NO Recombinant protein? NO 3. How much protein did you load? 34 ug/WELL (SEB-1 CELL LINE) AND 70 ug/WELL (HaCaT CELL LINE) I COULD NOT DETERMINE A DIFFERENCE BETWEEN THESE TWO CONCENTRATIONS ON THE BLOT How did you prepare the lysate for the analysis (protease inhibitors) SAMPLES WERE PREPARED USING NE-PER NUCLEAR AND CYTOPLASMIC EXTRACTION REAGENTS KIT FROM PIERCE AS DIRECTED IN THE MANUFACTURERS PROTOCOL. PROTEASE INHIBITORS WERE ADDED. Did you heat the samples? SAMPLES WERE HEATED TO 70º C FOR 10 MINUTES PRIOR TO LOADING ON GEL. 4. Primary Antibody ABCAM PPAR DELTA AB 8937 Specification (in which species was it raised against)? RABBIT POLYCLONAL At what dilution(s) have you tested this antibody? 1:1000 Incubation time, wash step? INCUBATED OVERNIGHT AT 4º C AND THEN WASHED 3 x 5 MINUTES IN TBS-T (0.05% TWEEN) 5. Secondary Antibody ANTI-RABBIT HRP FROM CELL SIGNALLING TECHNOLOGY. Specification (in which species was it raised against)? RABBIT At what dilution(s) have you tested this antibody? 1:2000 Incubation time, wash step? INCUABTED FOR 1 HOUR AT ROOM TEMPERATURE IN 5% DRY MILK, TWEEN-20 (0.05%) AND THEN WASHED 2 X 5 MINUTES IN TBS-T AND 1 X 5 MINUTES IN TBS Do you know whether the problems you are experiencing come from the secondary? SECONDARY ANTIBODY HAS BEEN USED NUMEROUS TIMES WITHOUT A PROBLEM 6. What detection method are you using? SUPERSIGNAL WEST PICO CHEMILUMINENCENT SUBSTRATE KIT FROM PIERCE AS DIRECTED. 7. Background bands Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a â??No primaryâ?? control) YES AND LANE IS CLEAR OF BANDS Is the blocking step sufficient? (We recommend blocking the membrane by adding 20 ml of blocking buffer (5% non-fat dry milk, 0.1% Tween-20 in TBS). Incubate for 2 h at room temperature or overnight at 4°C with agitation) BLOCKING FOR 1 HOUR HAS ALWAYS BEEN SUFFICENT FOR US. Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps) WASHING 3 X 5 MINUTES AS IS STANDARD IN MOST PROTOCOLS At what size are the bands migrating? Could they be degradation products of your target? THE MAJOR BAND IN QUESTION IS THE ONE THAT MIGRATES AROUND 70-75 kDA, WHY IS THIS ONE MORE INTENSE THAN THE OTEHRS??? WHICH BAND IS THE CORRECT ONE FOR PPAR DELTA???????? Please provide an image of your blot (as an e-mail attachment, a faxed image is not sufficient) 8. Optimization attempts How many times have you tried the Western? NUMEROUS TIMES WITH OTHER COMPANY ANTIBODY AND NEVER GOTTEN RESULTS LIKE THIS. BLOT IS FREE OF BACKGROUND JUST BANDS IN WRONG PLACE. Do you obtain the same results every time e.g. are background bands always in the same place? N/A What steps have you altered? N/A 9. Did you apply positive and negative controls along with the samples? Please specify. NEGATIVE CONTROL WAS PRIMARY ANTIBODY ALONE. WE DID NOT RUN ANY SAMPLES AS POSITIVE OR NEGATIVE CONTROLS.

Read More
Answer

Thank you for the details that you have provided us with. This antibody has been characterized in Western blotting using 3T3 cell lysate, and we have not specifically tested it with human samples (although it is expected to cross-react due to sequence homology). So I would strongly suggest running a positive control, such as 3T3 lysate. We saw a band at approximately 53 kDa using ab8937 with 3T3 lysate. I was unfortunately unable to open the attachment that you sent, but to try to get reduce the number of multiple bands that you are seeing, you may want to decrease the concentrations of both the primary and secondary antibodies. However, in your case the one thing that I think you need to do is run a positive control. If you need further assistance, please contact us again.

Read More

Answer

Thank you for your enquiry regarding ab8937. Unfortunately we don't supply Elisa Kits for this antibody. In the future, if you cannot find what you are looking for within the Abcam catalogue, you can click on the link to "The World's Antibody Gateway". The World's Antibody Gateway is a free search engine service provided by Abcam to help you to quickly find the antibodies that you are looking for. It is a free-text search engine developed by the Abcam team so that it searches the catalogues of all online antibody companies (currently 249). Should you require any further information to that shown on the datasheets, please get in touch with us and we will gladly assist you.

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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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