Anti-Hsc70 抗体 (ab1427)

Reviews (2)Q&A (4)References (11)

製品の概要

  • 製品名

    Anti-Hsc70 antibody
    Hsc70 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to Hsc70

  • 由来種

    Rabbit

  • 特異性

    The antibody detects a 73 kDa protein, corresponding to the apparent molecular weight of Hsc70 on SDS-PAGE immunoblots. No reactivity with inducible Hsp70 is detected.

  • アプリケーション

    適用あり: WB, IHC-Pmore details

  • 種交差性

    交差種: Mouse, Rat, Hamster, Cow, Human, Pig, Monkey

  • 免疫原

    Synthetic peptide:

    EKLQGKINDEDKQKI

    , corresponding to amino acids 555-570 of Human Hsc70

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • 特記事項


    The 70 kDa heat shock cognate protein, Hsc70, is closely related to Hsp70 and is part of the Hsp70 family. The family contains a number of highly-related protein isoforms ranging in size from 66 kDa to 78 kDa. These proteins include both cognate members which are found within major intracellular compartments and highly inducible isoforms which appear to be predominantly cytoplasmic or nuclear in distribution. Members of the Hsp70 family are molecular chaperones which are involved in many cellular functions such as protein folding, transport, maturation and degradation and they exert their function in an AATP-dependent manner.

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

  • バッファー

    pH: 7.20
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 50% Glycerol, 1% BSA
  • Concentration information loading...

  • 一次抗体 備考

    The 70 kDa heat shock cognate protein, Hsc70, is closely related to Hsp70 and is part of the Hsp70 family. The family contains a number of highly-related protein isoforms ranging in size from 66 kDa to 78 kDa. These proteins include both cognate members which are found within major intracellular compartments and highly inducible isoforms which appear to be predominantly cytoplasmic or nuclear in distribution. Members of the Hsp70 family are molecular chaperones which are involved in many cellular functions such as protein folding, transport, maturation and degradation and they exert their function in an AATP-dependent manner.

  • ポリ/モノ

    ポリクローナル

  • アイソタイプ

    IgG

  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab1427 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. Predicted molecular weight: 71 kDa.
IHC-P Use at an assay dependent concentration.

ターゲット情報

  • 機能

    Acts as a repressor of transcriptional activation. Inhibits the transcriptional coactivator activity of CITED1 on Smad-mediated transcription. Chaperone. Isoform 2 may function as an endogenous inhibitory regulator of HSC70 by competing the co-chaperones.

  • 組織特異性

    Ubiquitous.

  • 配列類似性

    Belongs to the heat shock protein 70 family.

  • ドメイン

    The N-terminal 1-386 residues constitute the ATPase domain, while residues 387-646 form the peptide-binding domain.

  • 翻訳後修飾

    Phosphorylated upon DNA damage, probably by ATM or ATR.
    ISGylated.

  • 細胞内局在

    Cytoplasm. Melanosome. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Translocates rapidly from the cytoplasm to the nuclei, and especially to the nucleoli, upon heat shock. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • 2410008N15Rik antibody
    • Constitutive heat shock protein 70 antibody
    • Epididymis luminal protein 33 antibody
    • Epididymis secretory sperm binding protein Li 72p antibody
    • Heat shock 70 kDa protein 8 antibody
    • Heat shock 70kD protein 10 antibody
    • Heat shock 70kD protein 8 antibody
    • Heat shock 70kDa protein 8 antibody
    • Heat shock cognate 71 kDa protein antibody
    • Heat shock cognate protein 54 antibody
    • Heat shock cognate protein 71 kDa antibody
    • Heat shock protein 8 antibody
    • Heat shock protein A8 antibody
    • Heat shock protein family A (Hsp70) member 8 antibody
    • Heat-shock70-KD protein 10, formerly antibody
    • HEL 33 antibody
    • HEL S 72p antibody
    • HSC54 antibody
    • HSC71 antibody
    • Hsc73 antibody
    • HSP71 antibody
    • HSP73 antibody
    • HSP7C_HUMAN antibody
    • HSPA10 antibody
    • HSPA8 antibody
    • LAP1 antibody
    • Lipopolysaccharide associated protein 1 antibody
    • LPS associated protein 1 antibody
    • LPS associated protein antibody
    • MGC102007 antibody
    • MGC106514 antibody
    • MGC114311 antibody
    • MGC118485 antibody
    • MGC131511 antibody
    • MGC29929 antibody
    • N-myristoyltransferase inhibitor protein 71 antibody
    • NIP71 antibody
    see all

画像

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsc70 antibody (ab1427)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsc70 antibody (ab1427)
    Ab1427 staining human normal testis tissue. Staining is localised to cytoplasmic and nuclear compartments.
    Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplif
  • Western blot - Anti-Hsc70 antibody (ab1427)
    Western blot - Anti-Hsc70 antibody (ab1427)This image is courtesy of an anonymous Abreview.
    All lanes : Anti-Hsc70 antibody (ab1427) at 1/2500 dilution

    Lane 1 : U2OS cell lysate
    Lane 2 : HeLa cell lysate

    Secondary
    All lanes : HRP-conjugated Goat polyclonal to Rabbit at 1/2000 dilution

    Performed under reducing conditions.

    Predicted band size: 71 kDa
    Observed band size: 71 kDa


    Exposure time: 2 minutes

    See Abreview

参考文献

This product has been referenced in:

  • Cao Y  et al. Augmentation of vaccine-induced humoral and cellular immunity by a physical radiofrequency adjuvant. Nat Commun 9:3695 (2018). Read more (PubMed: 30209303) »
  • Xu CY  et al. DJ-1 Inhibits a-Synuclein Aggregation by Regulating Chaperone-Mediated Autophagy. Front Aging Neurosci 9:308 (2017). Read more (PubMed: 29021755) »
See all 11 Publications for this product

ab1427 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

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1-6 of 6 Abreviews or Q&A

Western blot abreview for Anti-Hsc70 antibody

 Good
Abreviews
abreview image
Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing
Sample
Sheep Serum (Seminal Plasma)
Specification
Seminal Plasma
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More

Mr. Guillaume Tsikis

Verified customer

投稿 Jul 15 2013

Western blot abreview for Anti-Hsc70 antibody

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS and HeLa cells)
Loading amount
30 µg
Specification
U2OS and HeLa cells
Gel Running Conditions
Reduced Denaturing (12% Tris Glycine)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

投稿 Jan 18 2011

Question

BATCH NUMBER 202129 ORDER NUMBER 182689 DESCRIPTION OF THE PROBLEM no band SAMPLE 3T3 cell lysate PRIMARY ANTIBODY ab1427 1µg/ml in blocker for 1-2 hours, wash 3x5min in PBS DETECTION METHOD Fluorescence on a kodak IS4000MM using proper excitation and emission filters for the secondary antibodies POSITIVE AND NEGATIVE CONTROLS USED all samples were positive controls. ANTIBODY STORAGE CONDITIONS -20 degrees SAMPLE PREPARATION reduced and boiled 5min @95 degrees AMOUNT OF PROTEIN LOADED 20µg ELECTROPHORESIS/GEL CONDITIONS reducing 4-12% bis-tris gel 125V for 2 hours TRANSFER AND BLOCKING CONDITIONS overnight at 30V 5% milk + PBS blocker SECONDARY ANTIBODY Goat anti-rabbit Qdot 705 and Goat Anti-rabbit Alexa 750 at 1:1000 in blocker for 1 hour, wash 3x5min in PBS HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Changed the cell lysate to A431 cell lysate, no change

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Answer

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. I have read your technical questionnaire and I have a few comments. You are largely following an approach that I would recommend; using 3T3 mouse cells, a cell type that has been routinely used in the literature as a positive control for the detection of Hsc70 by western blotting. Can you tell me whether you have performed a ponceau red detection of the transfered protein or confirmed the integrity of the lysates using an alternative antibody that targets a "housekeeping" protein such as GAPDH. In the lab at Abcam we frequently find that a change to the blocking agent and/or antibody diluent produces more specific sensitive results. On this occasion I would like to recommend that you try applying the antibody using 3% BSA as a blocking agent, replacing milk. I would also like to recommend the use of TBST (as opposed to PBS) as the antibody diluent and wash buffer. Should a change to the blocking agent not improve your results please get back in touch with me and I will be happy to offer you a replacement vial (provided that the purchase was made within the past 90 days). I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Question

Is this antibody a purified antibody? If so, to what extent has it been purified?

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Answer

Dear Dr. Lloyd, Thank you for your enquiry. Regarding ab1427 (Hsc70 antibody), this product has been Protein A/G purified.

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Question

Customer is using this antibody in WB with recombinant protein but is not seeing a signal. The primary was used at 1-10 ug/ml and the signal detected was extremely weak. Lot# 132030.

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Answer

Thank you for your phone call and I'm sorry to hear that you are experiencing difficulty with ab1427. We have not received any other complaints regarding this antibody and it may be that the vial you received was damaged. I would be happy to send you a free of charge replacement vial or can have a credit note or refund issued to you. Please let me know how you would like to proceed and I look forward to hearing from you.

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Question

To what part of the Hsc70 molecule (N-end, C-end, middle, sequence) is the antibody directed?

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Answer

Thank you for your enquiry regarding ab1427. Synthetic peptide surrounding amino acid 559 of human Hsc70 was used to raise this antibody.

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