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使用敲除细胞株进行验证重组RabMAb

重组Anti-Hsp27抗体[EPR23938-256] - BSA and Azide free (Capture) (ab281198)

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Immunocytochemistry/ Immunofluorescence - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198)
  • Western blot - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198)
  • Western blot - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198)
  • Sandwich ELISA - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198)
  • Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23938-256] to Hsp27 - BSA and Azide free (Capture)
  • Suitable for: ICC/IF, Sandwich ELISA, WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free Unconjugated

概述

  • 产品名称

    Anti-Hsp27抗体[EPR23938-256] - BSA and Azide free (Capture)
    参阅全部 Hsp27 一抗

  • 描述

    兔单克隆抗体[EPR23938-256] to Hsp27 - BSA and Azide free (Capture)

  • 宿主

    Rabbit

  • 经测试应用

    适用于: ICC/IF, Sandwich ELISA, WBmore details

  • 种属反应性

    与反应: Human

  • 免疫原

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 常规说明

    Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198) is a BSA and Azide Free antibody supplied in an unconjugated format and it is suitable for sandwich ELISAs to quantify human Hsp27.

    The recommended pair for sandwich ELISA is:

    Capture: ab281198, human Hsp27 Capture Antibody (unconjugated)

    Detector: ab308543, human Hsp27 Detector Antibody (unconjugated)

    The recommended antibody orientation is based on internal optimization for ELISA-based assays. Antibody orientation is assay dependent and needs to be optimized for each assay type. Please note that the range provided for this antibody is only an estimation based on the performance of the product using the recommended antibody pair. Performance of the antibody pair will depend on the specific characteristics of your assay. We guarantee the product works in sandwich ELISA, but we do not guarantee the sensitivity or dynamic range of the antibody in your assay.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency. The antibodies are provided at an approximate concentration of 1 mg/ml as measured by the protein A280 method. Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    The recommended antibody orientation is based on internal optimization for ELISA-based assays. Antibody orientation is assay dependent and needs to be optimized for each assay type. Please note that the range provided for this antibody is only an estimation based on the performance of the product using the recommended antibody pair. Performance of the antibody pair will depend on the specific characteristics of your assay. We guarantee the product works in sandwich ELISA, but we do not guarantee the sensitivity or dynamic range of the antibody in your assay.

    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.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab281198于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
ICC/IF
1/100.
Sandwich ELISA
Use at an assay dependent concentration. Can be paired for Sandwich ELISA with PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918) and Streptavidin Conjugation Kit - Lightning-Link® (ab102921) and Biotinylation Kit / Biotin Conjugation Kit (Fast and Type A) - Lightning-Link® (ab201795) and Rabbit monoclonal [EPR23938-91] to Hsp27 - BSA and Azide free (Detector) (ab281048) and Rabbit monoclonal [EPR7278-114] to Hsp27 (phospho S82) - BSA and Azide free (Detector) (ab308543).
WB
1/1000. Predicted molecular weight: 23 kDa.
说明
ICC/IF
1/100.
Sandwich ELISA
Use at an assay dependent concentration. Can be paired for Sandwich ELISA with PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918) and Streptavidin Conjugation Kit - Lightning-Link® (ab102921) and Biotinylation Kit / Biotin Conjugation Kit (Fast and Type A) - Lightning-Link® (ab201795) and Rabbit monoclonal [EPR23938-91] to Hsp27 - BSA and Azide free (Detector) (ab281048) and Rabbit monoclonal [EPR7278-114] to Hsp27 (phospho S82) - BSA and Azide free (Detector) (ab308543).
WB
1/1000. Predicted molecular weight: 23 kDa.

靶标

  • 功能

    Involved in stress resistance and actin organization.

  • 组织特异性

    Detected in all tissues tested: skeletal muscle, heart, aorta, large intestine, small intestine, stomach, esophagus, bladder, adrenal gland, thyroid, pancreas, testis, adipose tissue, kidney, liver, spleen, cerebral cortex, blood serum and cerebrospinal fluid. Highest levels are found in the heart and in tissues composed of striated and smooth muscle.

  • 疾病相关

    Defects in HSPB1 are the cause of Charcot-Marie-Tooth disease type 2F (CMT2F) [MIM:606595]. CMT2F is a form of Charcot-Marie-Tooth disease, the most common inherited disorder of the peripheral nervous system. Charcot-Marie-Tooth disease is classified in two main groups on the basis of electrophysiologic properties and histopathology: primary peripheral demyelinating neuropathy or CMT1, and primary peripheral axonal neuropathy or CMT2. Neuropathies of the CMT2 group are characterized by signs of axonal regeneration in the absence of obvious myelin alterations, normal or slightly reduced nerve conduction velocities, and progressive distal muscle weakness and atrophy. Nerve conduction velocities are normal or slightly reduced. CMT2F onset is between 15 and 25 years with muscle weakness and atrophy usually beginning in feet and legs (peroneal distribution). Upper limb involvement occurs later. CMT2F inheritance is autosomal dominant.
    Defects in HSPB1 are a cause of distal hereditary motor neuronopathy type 2B (HMN2B) [MIM:608634]. Distal hereditary motor neuronopathies constitute a heterogeneous group of neuromuscular disorders caused by selective impairment of motor neurons in the anterior horn of the spinal cord, without sensory deficit in the posterior horn. The overall clinical picture consists of a classical distal muscular atrophy syndrome in the legs without clinical sensory loss. The disease starts with weakness and wasting of distal muscles of the anterior tibial and peroneal compartments of the legs. Later on, weakness and atrophy may expand to the proximal muscles of the lower limbs and/or to the distal upper limbs.

  • 序列相似性

    Belongs to the small heat shock protein (HSP20) family.

  • 翻译后修饰

    Phosphorylated in MCF-7 cells on exposure to protein kinase C activators and heat shock.

  • 细胞定位

    Cytoplasm. Nucleus. Cytoplasm > cytoskeleton > spindle. Cytoplasmic in interphase cells. Colocalizes with mitotic spindles in mitotic cells. Translocates to the nucleus during heat shock and resides in sub-nuclear structures known as SC35 speckles or nuclear splicing speckles.
  • Information by UniProt

  • 数据库链接

    • 别名

      • Heat shock 27kDa protein antibody
      • 28 kDa heat shock protein antibody
      • CMT2F antibody
      • DKFZp586P1322 antibody
      • epididymis secretory protein Li 102 antibody
      • Estrogen regulated 24 kDa protein antibody
      • Estrogen-regulated 24 kDa protein antibody
      • Heat shock 25kDa protein 1 antibody
      • Heat shock 27 kDa protein antibody
      • Heat shock 27kD protein 1 antibody
      • Heat shock 27kDa protein 1 antibody
      • Heat shock 28kDa protein 1 antibody
      • Heat Shock Protein 27 antibody
      • Heat shock protein beta 1 antibody
      • Heat shock protein beta-1 antibody
      • heat shock protein family B (small) member 1 antibody
      • HEL-S-102 antibody
      • HMN2B antibody
      • HS.76067 antibody
      • Hsp 25 antibody
      • HSP 27 antibody
      • Hsp 28 antibody
      • Hsp B1 antibody
      • Hsp25 antibody
      • HSP27 antibody
      • Hsp28 antibody
      • HspB1 antibody
      • HSPB1_HUMAN antibody
      • SRP27 antibody
      • Stress responsive protein 27 antibody
      • Stress-responsive protein 27 antibody
      see all

    图片

    • Immunocytochemistry/ Immunofluorescence - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198)
      Immunocytochemistry/ Immunofluorescence - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198)
      Immunocytochemistry/ Immunofluorescence analysis of wild-type and HSPB1 KO HAP1 (human chronic myelogenous leukemia near-haploid cell) labeling Hsp27 with ab281198 at 1/100 dilution followed by ab150081, secondary antibody at 1/1000 dilution. Confocal image showing cytoplasmic staining in wild-type HAP1 cells, and showing no staining in HSPB1 knockout HAP1 cells (shown in green). The counterstain antibody was ab195889 at 1/200 and was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100.
      Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
    • Western blot - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198)
      Western blot - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198)
      All lanes : Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198) at 1/1000 dilution

      Lane 1 : Human cerebellum tissue lysate
      Lane 2 : Human heart tissue lysate
      Lane 3 : Human liver tissue lysate
      Lane 4 : Human spleen tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 23 kDa
      Observed band size: 23-27 kDa why is the actual band size different from the predicted?


      Exposure time: 37 seconds


      Loading control: Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1/200000) (36KDa).
      Blocking/Diluting buffer and concentration 5% NFDM/TBST
      Low expression: cerebellum and spleen

    • Western blot - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198)
      Western blot - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198)
      All lanes : Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198) at 1/1000 dilution

      Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 23 kDa
      Observed band size: 27 kDa why is the actual band size different from the predicted?


      Exposure time: 180 seconds


      Loading control: Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1/200000) (36KDa).
      Blocking/Diluting buffer and concentration 5% NFDM/TBST

      Negative control: HT-1080.

    • Sandwich ELISA - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198)
      Sandwich ELISA - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198)
      Sandwich ELISA of ab308542 with the capture antibody dilution at 2 µg/mL and detector antibody dilution at 0.5 µg/mL. A549 + Anisomycin: A549 treated with 1 µg/ml anisomycin for 30 minutes, whole cell lysate. Serial dilutions of lysates were analyzed. A549 Control: untreated A549, whole cell lysate. Serial dilutions of lysates were analyzed.
    • Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198)
      Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198)

    实验方案

    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

    文献 (0)

    发表研究结果有使用 ab281198？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab281198 尚未被引用在任何文献中。

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