Anti-Lactate Dehydrogenase B抗体(ab75167)

概述

  • 产品名称Anti-Lactate Dehydrogenase B抗体
    参阅全部 Lactate Dehydrogenase B 一抗
  • 描述
    兔多克隆抗体to Lactate Dehydrogenase B
  • 经测试应用适用于: WB, ELISA, ICC/IF, IP, IHC-Pmore details
  • 种属反应性
    与反应: Human, Zebrafish
    预测可用于: Mouse, Rat, Rabbit, Horse, Chicken, Guinea pig, Cow, Cat, Dog, Pig, Chimpanzee, Spider Monkey
  • 免疫原

    A synthetic peptide corresponding to a region of Human lactate dehydrogenase B

  • 阳性对照
    • Fetal muscle lysate This antibody gave a positive result in IF/ICC when used in the following methanol fixed cell lines: DU145.

性能

应用

Our Abpromise guarantee covers the use of ab75167 in the following tested applications.

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

应用 Ab评论 说明
WB Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
ELISA Use at an assay dependent concentration. Titre using peptide based assay 1:62500
ICC/IF Use a concentration of 1 µg/ml.
IP Use a concentration of 5 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.

靶标

Anti-Lactate Dehydrogenase B antibody 图像



  • Predicted band size : 37 kDa
    Western blot analysis of MiaPaca-2 (lane 1) Panc-1 (lane 2) cell lysates (30µg/lane) labeling Lactate dehydrogenase B with ab75167 at 1:1000. A HRP-conjugated Goat anti-rabbit (1:4000) was used as the secondary antibody.
  • Lactate Dehydrogenase B was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Lactate Dehydrogenase B 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 ab75167.

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

    Band: 34kDa; Lactate Dehydrogenase B

  • ab75167 at 5 µg/ml staining lactate dehydrogenase B in Human placenta by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
  • Anti-Lactate Dehydrogenase B antibody (ab75167) at 1 µg/ml + fetal muscle lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size : 37 kDa
    Observed band size : 37 kDa
    Gel concentration: 12%
  • IHC image of ab75167 staining in human normal liver (bile duct cells) 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 ab75167, 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.

  • ab75167 at 5 µg/ml staining lactate dehydrogenase B in Human placenta by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
  • All lanes : Anti-Lactate Dehydrogenase B antibody (ab75167) at 1 µg/ml

    Lane 1 : Marker
    Lane 2 : Zebrafish brain homogenate at 20 µg
    Lane 3 : Zebrafish heart homogenate at 20 µg
    Lane 4 : Zebrafish liver homogenate at 20 µg
    Lane 5 : Zebrafish skeletal muscle homogenate at 20 µg

    Secondary
    Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 37 kDa
    Observed band size : 37 kDa


    Exposure time : 5 minutes
  • ICC/IF image of ab75167 stained DU145 cells. The cells were 100% methanol fixed (5 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 ab75167 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 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.

  • Application: IHC. Species+tissue/cell type: Human lung adenocarcinoma cell line A549. Primary antibody dilution: 1:100. Secondary antibody: Goat anti-rabbit AlexaFluor 488. Secondary antibody dilution: 1:400

Anti-Lactate Dehydrogenase B antibody (ab75167)参考文献

This product has been referenced in:
  • Usoskin D  et al. Unbiased classification of sensory neuron types by large-scale single-cell RNA sequencing. Nat Neurosci 18:145-53 (2015). Read more (PubMed: 25420068) »
  • 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). IHC ; Human . Read more (PubMed: 25973606) »

See all 3 Publications for this product

Product Wall

Application Western blot
Sample Mouse Tissue lysate - whole (Brain, heart, liver, muscle, retina)
Gel Running Conditions Reduced Denaturing (12% gel)
Loading amount 20 µg
Specification Brain, heart, liver, muscle, retina
Blocking step Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Mr. Connor Jankowski

Verified customer

提交于 Sep 08 2016

Application Western blot
Sample Cow Purified protein (heart)
Gel Running Conditions Reduced Denaturing (12%)
Loading amount 100 µg
Specification heart
Blocking step Milk as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Amber Howerton

Verified customer

提交于 May 18 2016

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