Anti-Lactate Dehydrogenase 抗体 (ab47010)

製品の概要

  • 製品名Anti-Lactate Dehydrogenase antibody
    Lactate Dehydrogenase 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Lactate Dehydrogenase
  • 特異性ab47010 cross-reacts with LDHA, B and C.
  • アプリケーション適用あり: WB, IP, ICC/IF, IHC-Pmore details
  • 種交差性
    交差種: Human, Zebrafish
    交差が予測される動物種: Mouse, Rat, Rabbit, Chimpanzee, Chinese Hamster
  • 免疫原

    Synthetic peptide corresponding to Human Lactate Dehydrogenase aa 300 to the C-terminus conjugated to Keyhole Limpet Haemocyanin (KLH).
    (Peptide available as ab47009, ab47009)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) A431 (Human epithelial carcinoma cell line) HEK293 (Human embryonic kidney cell line)

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab47010 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
WB Use a concentration of 1 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 37 kDa).Can be blocked with Human Lactate Dehydrogenase peptide (ab47009).
IP Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ターゲット情報

Anti-Lactate Dehydrogenase antibody 画像

  • All lanes : Anti-Lactate Dehydrogenase antibody (ab47010)

    Lane 1 : LDHA recombinant
    Lane 2 : LDHB recombinant
    Lane 3 : LDHC recombinant
    Lane 4 : HeLa


    Predicted band size : 37 kDa


    Exposure time : 30 seconds

    Observed bands

    Lane 1 :40 kDa

    Lane 2:37 kDa

    Lane 3:62 kDa

    Lane 4: 37 kDa

  • LDHA was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to LDHA 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, Hela whole cell extract 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 ab47010.

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

    Band: 36kDa; LDHA

  • All lanes : LDHA antibody (ab47010) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 37 kDa
    Observed band size : 36 kDa (why is the actual band size different from the predicted?)
  • ICC/IF image of ab47010 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab47010, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).
  • IHC image of Lactate Dehydrogenase staining in human breast carcinoma FFPE 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 ab47010, 1µg/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.
  • ab47010 at a 1/400 dilution staining LDHA in human skeletal muscle sections by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections) incubated for 20 minutes at 25°C. Heat mediated antigen retrieval used, EDTA ph 9.0, 100°C for 20 minutes. Secondary used undiluted, goat anti-mouse/rabbit IgG conjugated to HRP polymer.

    See Abreview

  • Anti-Lactate Dehydrogenase antibody (ab47010) at 1 µg/ml + Recombinant human LDHA protein (His tag) (ab93699) at 0.1 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 37 kDa


    Exposure time : 10 seconds

Anti-Lactate Dehydrogenase antibody (ab47010) 使用論文

This product has been referenced in:
  • Du Y  et al. Lysine malonylation is elevated in type 2 diabetic mouse models and enriched in metabolic associated proteins. Mol Cell Proteomics 14:227-36 (2015). Read more (PubMed: 25418362) »
  • Sun W  et al. Lactate dehydrogenase B is associated with the response to neoadjuvant chemotherapy in oral squamous cell carcinoma. PLoS One 10:e0125976 (2015). Human . Read more (PubMed: 25973606) »

See all 7 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Cow Cell lysate - whole cell (VVEC)
Gel Running Conditions Reduced Denaturing (12%)
Loading amount 20 µg
Treatment ATP 100 uM TC
Specification VVEC
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
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投稿 Jun 21 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (lung)
Antigen retrieval step None
Permeabilization No
Specification lung
Blocking step peroxidase as blocking agent for 5 minute(s) · Concentration: 0.03%
Fixative Formaldehyde
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投稿 Nov 18 2015

Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Liver)
Permeabilization No
Specification Liver
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative Acetone
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投稿 Apr 16 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (SH-SY5Y cells)
Specification SH-SY5Y cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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投稿 Jan 27 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Non-reduced Denaturing (10%)
Sample Human Cell lysate - whole cell (A 549 cells)
Specification A 549 cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Dr. Christine Vohwinkel

Verified customer

投稿 Nov 18 2014

The antibody ab85319, anti-LDHb, reacts with western blots of recombinant LDHb but not blots containing recombinant LDHa. Please let us know if you have any questions.

As my colleague mentioned in a previous message, ab4710 is not an antibod...

Read More

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....

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Zebrafish Cell lysate - whole cell (zebrafish embryo - 14dpf)
Loading amount 10 µg
Specification zebrafish embryo - 14dpf
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

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投稿 Oct 21 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (skeletal muscle)
Specification skeletal muscle
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: EDTA pH 9, 100C, 20 minutes
Permeabilization No
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投稿 Dec 10 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"