Overview

  • Product name
  • Description
    Rabbit polyclonal to ALDH1A3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human, African green monkey, Syrian hamster
    Predicted to work with: Rabbit, Horse, Cow, Chimpanzee, Macaque monkey, Gorilla, Chinese hamster, Orangutan
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human ALDH1A3.

    Read Abcam's proprietary immunogen policy

  • Positive control
    • This antibody gave a positive signal in Human, Mouse and Rat Kidney tissue lysates. IHC-P: human stomach tissue sections. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: PC3

Properties

Applications

Our Abpromise guarantee covers the use of ab129815 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
IP Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

Images

  • All lanes : Anti-ALDH1A3 antibody (ab129815) at 1 µg/ml

    Lane 1 : Human kidney tissue lysate - total protein (ab30203)
    Lane 2 : Kidney (Mouse) Tissue Lysate
    Lane 3 : Kidney (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 56 kDa
    Observed band size: 56 kDa
    Additional bands at: 12 kDa, 32 kDa, 37 kDa, 47 kDa, 50 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 1 minute


    This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab129815 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
  • IHC image of ab129815 staining in human stomach formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab129815, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • ab129815 stained PC3 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 ab129815 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 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.

  • ALDH1A3 was immunoprecipitated using 0.5mg Mouse Kidney tissue lysate, 5µg of Rabbit polyclonal to ALDH1A3 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Mouse Kidney tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab129815.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

    Band: 56kDa; ALDH1A3

References

This product has been referenced in:
  • Chen L  et al. Everolimus Reverses Palbociclib Resistance in ER+ Human Breast Cancer Cells by Inhibiting Phosphatidylinositol 3-Kinase(PI3K)/Akt/Mammalian Target of Rapamycin (mTOR) Pathway. Med Sci Monit 25:77-86 (2019). Read more (PubMed: 30605443) »
  • Ryu WJ  et al. A Therapeutic Strategy for Chemotherapy-Resistant Gastric Cancer via Destabilization of Both ß-Catenin and RAS. Cancers (Basel) 11:N/A (2019). Read more (PubMed: 30965636) »
See all 7 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (4-12% gradient gel Bis-Tris)
Sample
Mouse Tissue lysate - whole (Intestine)
Specification
Intestine
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 23 2014

Application
Western blot
Loading amount
100 µg
Gel Running Conditions
Reduced Denaturing (12)
Sample
Dog Cell lysate - whole cell (MDCK cells)
Specification
MDCK cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 18 2013

Application
Western blot
Loading amount
100 µg
Gel Running Conditions
Reduced Denaturing (12)
Sample
Syrian Hamster Cell lysate - whole cell (BHK cells)
Specification
BHK cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 18 2013

Application
Western blot
Loading amount
100 µg
Gel Running Conditions
Reduced Denaturing (12)
Sample
African Green Monkey Cell lysate - whole cell (COS cells)
Specification
COS cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 18 2013

Application
Western blot
Loading amount
100 µg
Gel Running Conditions
Reduced Denaturing (12)
Sample
Rat Cell lysate - whole cell (NRK cells)
Specification
NRK cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 18 2013

Application
Western blot
Loading amount
100 µg
Gel Running Conditions
Reduced Denaturing (12)
Sample
Human Cell lysate - whole cell (HEK293F cells)
Specification
HEK293F cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 18 2013

Application
Western blot
Loading amount
100 µg
Gel Running Conditions
Reduced Denaturing (12)
Sample
Mouse Cell lysate - whole cell (NIH/3T3 cells)
Specification
NIH/3T3 cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Oct 18 2013

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