Overview

  • Product name
    Anti-Lactate Dehydrogenase Isoenzyme V antibody
    See all Lactate Dehydrogenase Isoenzyme V primary antibodies
  • Description
    Sheep polyclonal to Lactate Dehydrogenase Isoenzyme V
  • Host species
    Sheep
  • Tested applications
    Suitable for: IHC-P, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Human Lactate Dehydrogenase V, purified from human placenta.

  • Positive control
    • This antibody gave a positive signal in the following lysates: Human Kidney Tissue; Human Liver Tissue; HEK293 Whole Cell; HepG2 Whole Cell.

Properties

Applications

Our Abpromise guarantee covers the use of ab9002 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
IHC-P Use at an assay dependent dilution. PubMed: 12942121
WB Use a concentration of 1 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
IHC-Fr 1/100. PubMed: 21569415Snap freeze tissue in liquid nitrogen and store. Then the sections should be fixed for 10 minutes in acetone and rehydrated in PBS.

Target

  • Relevance
    Lactate dehydrogenase 5 (LDH5) is one of five LDH isoenzymes and important for promoting anaerobic glycolysis. LDH5 is transcriptionally regulated by the hypoxia inducible factors (HIF) 1a and 2a.
  • Cellular localization
    Cytoplasmic
  • Database links
  • Alternative names
    • Cell proliferation-inducing gene 19 protein antibody
    • L lactate dehydrogenase A chain antibody
    • Lactate dehydrogenase A antibody
    • LDH A antibody
    • LDH M antibody
    • LDH muscle subunit antibody
    • LDH1 antibody
    • LDH5 antibody
    • LDHA antibody
    • PIG 19 antibody
    • PIG19 antibody
    • Renal carcinoma antigen NY-REN-59 antibody
    see all

Images

  • All lanes : Anti-Lactate Dehydrogenase Isoenzyme V antibody (ab9002) at 1 µg/ml

    Lane 1 : Human kidney tissue lysate - total protein (ab30203)
    Lane 2 : Human liver tissue lysate - total protein (ab29889)
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Rabbit polyclonal Secondary Antibody to Sheep IgG - H&L (HRP) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 36 kDa
    Observed band size: 36 kDa


    Exposure time: 30 seconds
  • Immunohistochemical analysis of Human head and neck cancer tissue, staining Lactate Dehydrogenase Isoenzyme V (red) with ab9002 at 1/100 dilution. Sections were fixed in acetone before being incubated with primary antibody overnight at 4°C. A Cy3-conjugated donkey anti-sheep IgG (1/600) was used as the secondary antibody.

References

This product has been referenced in:
  • Tseng PL  et al. Decreased succinate dehydrogenase B in human hepatocellular carcinoma accelerates tumor malignancy by inducing the Warburg effect. Sci Rep 8:3081 (2018). Read more (PubMed: 29449614) »
  • Koukourakis MI  et al. Normal tissue radioprotection by amifostine via Warburg-type effects. Sci Rep 6:30986 (2016). WB . Read more (PubMed: 27507219) »
See all 21 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Testis)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA Buffer pH 9.0
Permeabilization
Yes - Wash Buffer from Dako with Tween
Specification
Testis
Fixative
Paraformaldehyde

Mr. Rudolf Jung

Verified customer

Submitted Jul 19 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Bladder Tumor Cell Line)
Permeabilization
Yes - 01% v/v Triton X-100
Specification
Bladder Tumor Cell Line
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Dr. Dimitra Kalamida

Verified customer

Submitted Sep 12 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (prostate cancer cell line)
Gel Running Conditions
Reduced Denaturing (10%acrylamide)
Loading amount
20 µg
Specification
prostate cancer cell line
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Stamatia Pouliliou

Verified customer

Submitted Jun 26 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (Epithelial cells)
Gel Running Conditions
Non-reduced Denaturing (10)
Loading amount
30 µg
Specification
Epithelial cells
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Mr. Achilleas Mitrakas

Verified customer

Submitted Jun 15 2015

Answer

Thank you for your phone call and your interest in our products.
As discussed over the phone, the Anti-Lactate Dehydrogenase Isoenzyme V antibody (ab9002) has been used by several research groups in performing immunohistochemical staining. Having reviewed the data we have for this antibody I would suggest it should work well in the tissue you have. The target has been shown to be expressed especially in cancerous tissue, but if you are looking for a different positive control you could potentially use human muscle tissue as we have done using the anti-Lactate Dehydrogenase Isoenzyme V antibody ab101562.
As you have said, the antigen retrieval used in the reference by Koukourakis et al. (2003) does not explicitly say which antigen retrieval buffer they found to work the best. However, in a separate paper staining of Beclin 1 was performed using citrate buffer (pH 7.2) using the microwave (3x5 min) (Koukourakis et al. (2010)). I would therefore not be surprised if this was used with the anti-LDH5 as well. However, in order to achieve the best staining with your tissue I would suggest trying citrate buffer (pH 6) as well as Tris/EDTA (pH 9) to see which would provide the best results. Using the pressure cooker should not be a problem. again, I would suggest optimising the time used (initially using 3, 5 and 10 minutes).
Additionally, I have had a look at the Novolink detection substrate and I don’t think it is likely to be suitable as it seems it is designed for mouse or rabbit primary antibodies. Ab9002 is a sheep polyclonal and would therefore need a secondary antibody directed against sheep IgG to work. If you would prefer to use a rabbit antibody you could look at ab101562 which should also work in the experiment you are planning.
I hope this information has been of help. If you have any further questions, please do not hesitate to let me know.

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Question
Answer

Thank you for your phone call.

DISCOUNT CODE: xxx
Expiration date: xxx

I am very pleased to hear you would like to accept our offer and test ab9002 in mouse. This code will give you: 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for mouse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
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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: https://www.abcam.com/collaborationdiscount.

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Answer

Thank you for contacting Abcam. I’ve had a look into this and done a bit of a web search and as far as I can see LDH5, LDHM and LDHA are all different names for the same protein, as stated in this paper, http://www.pnas.org/content/107/5/2037.long. Therefore, ab47010 can be used to detect LDH5.  I hope this information is helpful.  Please do not hesitate to contact us if you have any additional questions.  

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Question
Answer

From our testing we would expect our anti LDH-V to show a single band of approximately 36 kDa by Western blot - this band corresponds to the A subunit (sometimes called the muscle subunit) which LDH-V is entirely composed of.

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