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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)

Reviews (31)Q&A (13)References (223)
Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
  • Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
  • Immunocytochemistry/ Immunofluorescence - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
  • Flow Cytometry (Intracellular) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
  • ELISA - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
  • Immunocytochemistry/ Immunofluorescence - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
  • Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
  • Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
  • Immunoprecipitation - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
  • Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
  • Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
  • Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
  • Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y92] to PDGFR alpha + PDGFR beta - C-terminal
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP, ELISA
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 405 Alexa Fluor® 488 Alexa Fluor® 594 Carrier Free HRP PE

Overview

  • Product name

    Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal

  • Description

    Rabbit monoclonal [Y92] to PDGFR alpha + PDGFR beta - C-terminal

  • Host species

    Rabbit

  • Specificity

    Expression levels of the target protein vary with sample type and some optimisation may be required.

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP, ELISAmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human PDGFR beta aa 1050 to the C-terminus. The exact sequence is proprietary.
    Database link: P09619
    (Peptide available as ab263451)

  • Positive control

    • WB: N-GST tagged Human PDGF Receptor beta (aa557 to 1106) recombinant protein, N-GST tagged Human PDGF Receptor alpha (aa550 to 1089) recombinant protein, NIH/3T3 cell lysate. SH-SY5Y cell lysate. Rat brain and heart tissue lysate. Mouse brain tissue lysate. Human fetal brain tissue lysate. Human skeletal muscle tissue lysate. ICC/IF: NIH/3T3 cells. IHC-P: Human prostatic carcinoma, lung cancer, breast and spleen tissue. Flow Cyt (intra): NIH/3T3 cells. IP: NIH/3T3 cell lysate.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32570 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
Flow Cyt (Intra)
1/20.
WB (4)
1/5000 - 1/20000. Predicted molecular weight: 124 kDa.

For samples expressing low levels of PDGFR beta, the amount of lysate loaded may need to be increased to allow detection. 
IHC-P (8)
1/50 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Optimisation of the IHC protocol may be required depending on the sample used.
ICC/IF (8)
1/100.
IP
1/20.
ELISA
Use at an assay dependent concentration.

Primary antibody concentration range: 1000 - 0 ng/mL
Notes
Flow Cyt (Intra)
1/20.
WB
1/5000 - 1/20000. Predicted molecular weight: 124 kDa.

For samples expressing low levels of PDGFR beta, the amount of lysate loaded may need to be increased to allow detection. 
IHC-P
1/50 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Optimisation of the IHC protocol may be required depending on the sample used.
ICC/IF
1/100.
IP
1/20.
ELISA
Use at an assay dependent concentration.

Primary antibody concentration range: 1000 - 0 ng/mL

Target

Images

  • Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    All lanes : Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/1000 dilution

    Lane 1 : Recombinant human PDGFR beta protein (ab60833)
    Lane 2 : Recombinant human PDGFR alpha protein (ab84797)

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 124 kDa



    Blocking/Diluting buffer and concentration: 5% NFDM/TBST

    Exposure time: 3 seconds

  • Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    All lanes : Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/5000 dilution

    Lane 1 : Wild-type SH-SY5Y cell lysate
    Lane 2 : PDGFR beta knockout SH-SY5Y cell lysate
    Lane 3 : Human Skeletal Muscle tissue lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 30 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 124 kDa
    Observed band size: 170 kDa why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab32570 observed at 170 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab32570 was shown to react with PDGFRB in wild-type SH-SY5Y cells in Western blot with weakened signal observed in PDGFRB knockout cell line ab273749 (knockout cell lysate ab275523). Wild-type SH-SY5Y and PDGFRB knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab32570 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging. The weak signal observed in PDGFRB knockout cell line should be PDGFRA.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)

    ab32570 staining PDGFR beta in human lung cancer tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Shows positive staining on stromal cells. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Samples were incubated with primary antibody (1/500). An HRP-conjugated Goat anti-rabbit IgG (ready to use) was used as the secondary antibody. Counter stained with Hematoxylin.

  • Immunocytochemistry/ Immunofluorescence - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Immunocytochemistry/ Immunofluorescence - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)Image from Miyata M et al., J Biol Chem. 2009 Sep 4;284(36):24595-609. Epub 2009 Jul 9. Fig 1.; doi: 10.1074/jbc.M109.016436; September 4, 2009, The Journal of Biological Chemistry, 284, 24595-24609.

    Immunofluorescence analysis of NIH/3T3 (Mouse embryo fibroblast cell line) cells stimulated with PDGF, staining PDGFR beta with unpurified ab32570.

  • Flow Cytometry (Intracellular) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Flow Cytometry (Intracellular) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)

    Intracellular Flow Cytometry analysis of NIH/3T3 (Mouse embryo fibroblast cell line) cells labeling PDGFR beta (red) with ab32570 at a 1/20 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with primary and secondary antibodies. 

     

     

  • ELISA - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    ELISA - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)

    ELISA showing primary antibody ab32570 binding to the antigen Human PDGFR alpha protein and Human PDGFR beta protein. 

    Primary antibody concentration ranges from 0 - 1000 ng/mL. 

    Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) was used as a secondary antibody at 1/2500 dilution. 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)

    Immunohistochemical staining of paraffin embedded human spleen with purified ab32570 at a working dilution of 1/50. The secondary antibody used is ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Immunocytochemistry/ Immunofluorescence - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)

    Immunofluorescence staining of NIH/3T3 (Mouse embryo fibroblast cell line) cells with purified ab32570 at a working dilution of 1 in 100, counter-stained with DAPI. Tubulin was stained with mouse anti-tubulin at a dilution of 1/1000 (ab7291) and Alexa Fluor® 594 goat anti-mouse at a dilution of 1/500 (ab150120) . The secondary antibody was ab150077 Alexa Fluor® 488 goat anti rabbit, used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100.

    The negative controls are shown in the bottom middle and right hand panels - for the first negative control, purified ab32570 was used at a dilution of 1/200 followed by an Alexa Fluor® 555 goat anti-mouse antibody at a dilution of 1/500 and for the second negative control mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab15007) were used.

  • Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    All lanes : Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/10000 dilution (purified)

    Lane 1 : Rat brain tissue lysate
    Lane 2 : Rat heart tissue lysate
    Lane 3 : Mouse brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 124 kDa
    Observed band size: 175 kDa why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Different batches of ab32570 were tested on Rat brain lysate at 1.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 175 kDa.
  • Immunoprecipitation - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Immunoprecipitation - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)

    ab32570 (purified) at 1/20 immunoprecipitating PDGFR beta in NIH/3T3 (Mouse embryo fibroblast cell line) (Lane 1 and 2). Lane 3 - PBS.

    For western blotting a HRP-conjugated anti-rabbit IgG specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/10000 dilution (purified) + SH-SY5Y (Human neuroblastoma cell line from bone marrow) cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 124 kDa
    Observed band size: 175 kDa why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/5000 dilution (purified) + Human fetal brain tissue lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 124 kDa
    Observed band size: 175 kDa why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Western blot - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/50000 dilution (purified) + NIH/3T3 (Mouse embryo fibroblast cell line) cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 124 kDa
    Observed band size: 175 kDa why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)

    ab32570 staining PDGFR beta in human breast tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Shows positive staining on stromal cells. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Samples were incubated with primary antibody (1/500). An HRP-conjugated Goat anti-rabbit IgG (ready to use) was used as the secondary antibody. Counter stained with Hematoxylin.

  • Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)
    Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal (ab32570)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (223)

Publishing research using ab32570? Please let us know so that we can cite the reference in this datasheet.

ab32570 has been referenced in 223 publications.

  • Becerril C  et al. Mesenchymal-Epithelial Transition in Fibroblasts of Human Normal Lungs and Interstitial Lung Diseases. Biomolecules 11:N/A (2021). PubMed: 33806618
  • Miyauchi K  et al. Renal interstitial fibroblasts coproduce erythropoietin and renin under anaemic conditions. EBioMedicine 64:103209 (2021). PubMed: 33508746
  • Su F  et al. Progression of prostate carcinoma is promoted by adipose stromal cell-secreted CXCL12 signaling in prostate epithelium. NPJ Precis Oncol 5:26 (2021). PubMed: 33753872
  • Petre A  et al. A novel experimental approach to evaluate guided bone regeneration (GBR) in the rat femur using a 3D-printed CAD/CAM zirconia space-maintaining barrier. J Adv Res 28:221-229 (2021). PubMed: 33364058
  • Uzunalli G  et al. Structural disruption of the blood-brain barrier in repetitive primary blast injury. Fluids Barriers CNS 18:2 (2021). PubMed: 33413513
View all Publications for this product

Customer reviews and Q&As

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31-40 of 44 Abreviews or Q&A

Western blot abreview for Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Rat Cell lysate - whole cell (rat vascular smooth muscle cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
100000 cells
Specification
rat vascular smooth muscle cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 05 2013

Western blot abreview for Anti-PDGFR alpha + PDGFR beta antibody [Y92] - C-terminal

 Good
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (liver cancer)
Gel Running Conditions
Reduced Denaturing (8% gel)
Loading amount
15 µg
Specification
liver cancer
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 09 2012

Question

What sample and protocol was used for the failed flow cytometry test?

Read More
Answer

Permeabilized NIH/3T3 cells were used with ab32570 for the flow cytometry test. The protocol is at the following link on the Epitomics website:

http://www.epitomics.com/products/support/fc .

Read More

Question

Hello,

I require the concentrations for the following antibodies please?

·Ab32570 lot # GR7483-3

· Ab 92574 lot# GR37919-4

Thank you.

Read More
Answer

Thank you for contacting us.

The concentration for ab32570 lot GR7483-3 is XXX

Conc. for ab92574 lot GR37919-4 is XXXX

Please note these are rabbit monoclonals antibodies. They are very specific to their targets so the working dilutions will be much higher than polyclonal antibodies.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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

Read More

Question

Hello,

The correct lot number is GR67483-3. The concentration for this antibody was supplied to us on a different lot number, it is important that we get this information as I will be running a non-immune to match.

Kind Regards,

Read More
Answer

Thank you for confirming the correct batch number. This is to confirm that I have contacted the Lab to get the concentration values for you. Thank you for your patience in advance.

Read More

Question

Hello,

I require the concentrations for the following antibodies please?

·Ab32570 lot # GR7483-3

·Thank you.

Read More
Answer

Thank you for your enquiry.

This is a monoclonal rabbit antibody and we normally do not provide the concentration. However, we list the starting recommended dilutions factors for each tested application (please see the on-line datasheet):

ICC/IF: 1/100


IHC-P: 1/100

IP: 1/30

WB: 1/10,000

Rabbit monoclonal antibodies are more specific and have higher affinity than mouse monoclonals or rabbit polyclonal antibodies.You can find some further information if you visit the following site at:

https://www.abcam.com/index.html?pageconfig=resource&rid=15279

Would you be so kind to check your record again and provide the correct batch number for this product? Unfortunately, our system does not recognize GR7483-3.

If you need any further assistance in the future, please do not hesitate to contact me.

Read More

Question

Concentration of ab51037, ab32570 and ab92574?

Read More
Answer

Thank you for contacting us.
As discussed over the phone, could you please send me the lot numbers?
Thank you.

Read More

Question

Good afternoon,

I bought 2 antobodies from Abcam:

- Anti-NG2 (LHM 2) - ab20156
- Anti-PDGF receptor beta - ab32570

I need some informations.



Can you tell me if I need to permeabilize the cells?

Are these membrane antibodies?



Thank you

Patricia
--

Patricia Pitrez Pereira, PhD Student



MIT Portugal Bioengineering Systems
http://www.mitportugal.org/


Biocant - Center of Innovation and Biotechnology

Nucleo 4, Lote 3
3060-197 Cantanhede - Portugal

http://www.biocant.pt/
Phone: +351 231 419 040
Fax: +351 231 419 049
http://biomaterials.biocant.pt/

Read More
Answer

Thank you for your email and for your telephone enquiry earlier today.

As discussed on the telephone, PDGF receptor beta is a membrane protein, and so I would recommend that detection with ab35270 would not usuallyrequire permeablization. Though this may require some individual optimization.

NG2 is an apical cell membrane protein. Reviewing the details after our phone conversation,as this is a membrane protein, it will not require permeabilization. However, as this protein can alsorelocate to other parts of the cell, such as thelamellipodia, I can recommend you may require a gentle permeabilization step. A strong detergent like Triton may not be suitable, as this can disrupt membrane proteins and significantly affect the staining. I can suggest it would be more appropriate to include more gentle detergent such as 0.2% Tween or saponin in the antibody dilution buffers and wash buffers. This will help to keep the cells permeabilised. Again, this is a guideline only and may require some individual optimization.

Usually for membrane proteinswe would not recommend to use organic solventsto fix the samples, as this also permeabilizes the cells and can cause disruption to antibody epitopes on membrane proteins.However, I do notice that an Abreview on teh datasheet forthe NG2 antibody indicates that acetone fixation worked best in frozen sections. This does highlight and indicate that although we canprovide recommendations, individual optimization is sometimes required.

https://www.abcam.com/NG2-antibody-LHM-2-ab20156.html&tab=abreviews&intabreviewid=6408

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Question

Recently I order some antibodies from your company to detect NG2 (ab101679) and PDGFRb (ab32570).
It is supposed that it was reported that they worked for immunofluorescence. I have tried with free floating
sections, frozen sections and flow cytometry without any luck.
Do you know if there is a problem with the recent lot for these antibodies?
Also, I noticed that the anti-NG2 serum look like a precipitate and it looks opaque. Is that normal for the
anti-NG2 antiserum?
Even, I give a little aliquot to some one from a different lab that is testing antibodies from Biolegend and he did
not have luck detecting NG2+ or PDGFRb+ cells. And we tried to detect the signal in tissues were they have been
reported like brain and heart. I am very disappointed with these antibodies because I have spend almost 1000 dollars
and two weeks without any results.
Could you give me some advice or replace the antibodies for other clones that work in frozen sections like
NG2 (ab83508)?

Read More
Answer

Thank you for contacting us. I am sorry to hear that these antibodies have not been working in your applications. Can you please provide some additional detail about your experiments? If the antibodies are not working as stated on our datasheet and you have purchased them in the past six months, I will be happy to offer you free of charge replacements, or a credit or refund.

1) What type of results were you getting in IHC-Fr? Did you observe high background, no signal, or incorrect localization? It would be very helpful if you could please send an image of your results.

2) What species and sample type were you working with? How were the samples fixed and permeabilized?

3) What blocking agent was used?

4) How long were the primary and secondary antibodies incubated and what dilutions were used? It would be helpful if you could provide some details about the secondary antibody or the catalog number of the product used.

I will be happy to assist you further once I have this additional information.

Read More

Question

Thanks. Do you recommend using rabbit IgG1 as isotype control for this antibody during immunocytofluorescence staining?

Read More
Answer

Thank you for contacting us.

Yes, the rabbit IgG1 can be used as Isotype control.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

31-40 of 44 Abreviews or Q&A

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