製品の概要

  • 製品名Anti-Grp75 antibody [JG1]
    Grp75 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [JG1] to Grp75
  • アプリケーション適用あり: WB, IP, ICC/IF, IHC-P, Flow Cytmore details
  • 種交差性
    交差種: Mouse, Rat, Guinea pig, Hamster, Dog, Human, Non Human Primates
    交差が予測される動物種: Cow
  • 免疫原

    Synthetic peptide corresponding to Mouse Grp75 aa 661-679.
    Sequence:

    GSGSSGTGEQKEDQKEEKQ


    Database link: P38647

  • 特記事項

     

     

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab2799 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 1/500. Detects a band of approximately 74 kDa (predicted molecular weight: 74 kDa).
IP Use at an assay dependent concentration.
ICC/IF 1/50 - 1/200.

Staining of mtHSP 70 in DAP.3 cells results in a worm-like staining pattern, consistent with mitochondrial localization.

IHC-P Use a concentration of 1 µg/ml.
Flow Cyt 1/100.

ターゲット情報

  • 機能Implicated in the control of cell proliferation and cellular aging. May also act as a chaperone.
  • 配列類似性Belongs to the heat shock protein 70 family.
  • 翻訳後修飾Phosphorylated upon DNA damage, probably by ATM or ATR.
  • 細胞内局在Mitochondrion.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 75 kDa glucose regulated protein antibody
    • 75 kDa glucose-regulated protein antibody
    • CSA antibody
    • Glucose Regulated Protein antibody
    • Grp 75 antibody
    • GRP-75 antibody
    • GRP75 antibody
    • GRP75_HUMAN antibody
    • Heat shock 70 kDa protein 9 antibody
    • Heat shock 70kD protein 9 antibody
    • heat shock 70kDa protein 9 antibody
    • Heat shock 70kDa protein 9B antibody
    • Heat shock protein 74 kDa A antibody
    • Heat shock protein A antibody
    • Heat shock protein cognate 74 antibody
    • Hsc74 antibody
    • Hsp74 antibody
    • Hsp74a antibody
    • HSPA9 antibody
    • Hspa9a antibody
    • HSPA9B antibody
    • MGC4500 antibody
    • mitochondrial antibody
    • Mortalin 2 antibody
    • Mortalin antibody
    • Mortalin perinuclear antibody
    • Mortalin2 antibody
    • MOT 2 antibody
    • MOT antibody
    • MOT2 antibody
    • Mthsp70 antibody
    • p66 mortalin antibody
    • P66 MOT antibody
    • PBP74 antibody
    • Peptide binding protein 74 antibody
    • Peptide-binding protein 74 antibody
    • Stress 70 protein mitochondrial antibody
    • Stress 70 protein mitochondrial precursor antibody
    • Stress-70 protein antibody
    see all

Anti-Grp75 antibody [JG1] 画像

  • Immunocytochemistry/Immunofluorescence analysis of Grp75 (green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% BSA for 15 minutes at room temperature. Cells were incubated with (left panel) or without (right panel) ab2799 (1:50) for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat-anti-mouse IgG secondary antibody (1:400) for 30 minutes at room temperature. F-Actin (red) was stained with Dylight 554 phalloidin, and nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.

  • Immunohistochemistry was performed on normal biopsies of deparaffinized human testis tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer and microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a Mouse Monoclonal Antibody recognizing Grp75 (ab2799) or without primary antibody (negative control) overnight at 4�C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
  • ICC/IF image of ab2799 stained HeLa cells. The cells were 4% PFA fixed (10 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 (ab2799, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.

  • Anti-Grp75 antibody [JG1] (ab2799) at 1/750 dilution + Mouse brain tissue lysates at 10 µg

    Secondary
    HRP conjugated donkey polyclonal to mouse IgG3 at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 74 kDa
    Observed band size : 74 kDa


    Exposure time : 3 minutes

    This image is courtesy of an anonymous abreview.

    Primary incubated for 1 hour 30 minutes at 24°C.

  • Anti-Grp75 antibody [JG1] (ab2799) at 1/750 dilution

    Secondary
    HRP conjugated donkey anti mouse IgG3 at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 74 kDa
    Observed band size : 74 kDa


    Exposure time : 5 minutes

    This image is courtesy of an anonymous abreview.

    See Abreview

  • ab2799 (1µg/ml) staining GPR75 in human frontal cerebral cortex using an automated system (DAKO Autostainer Plus). Using this protocol there is strong mitochondrial staining.
    Insert depicts negative control (no primary antibody).
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min 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, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • Overlay histogram showing HepG2 cells stained with ab2799 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2799, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG3 [MG3-35] (ab18394, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Immunocytochemistry/Immunofluorescence analysis of human fibroblasts labelling Grp75 with ab2799.

Anti-Grp75 antibody [JG1] (ab2799) 使用論文

This product has been referenced in:
  • Ben Safta T  et al. Granzyme B-activated p53 interacts with Bcl-2 to promote cytotoxic lymphocyte-mediated apoptosis. J Immunol 194:418-28 (2015). ICC/IF ; Human . Read more (PubMed: 25404359) »
  • Sato J  et al. Family with Sequence Similarity 5, Member C (FAM5C) Increases Leukocyte Adhesion Molecules in Vascular Endothelial Cells: Implication in Vascular Inflammation. PLoS One 9:e107236 (2014). ICC/IF ; Human . Read more (PubMed: 25251368) »

See all 13 Publications for this product

Product Wall

Merci de nous avoir contactés. L'anticorps ab2799 est dirigé contre la protéine Grp75, référence SwissProt http://www.uniprot.org/uniprot/P38646 La liste des anti-Hsp70 (SwissProt http://www.uniprot.org/uniprot/P08107 ) est : http://ww...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - other (EL4)
Loading amount 15 µg
Specification EL4
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

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Verified customer

投稿 Sep 06 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Brain)
Specification Brain
Fixative microwave euthanasia
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Trilogy
Permeabilization Yes - tween20 (0.05%)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Username

Mr. Adrian Epstein

Verified customer

投稿 Aug 02 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - other (HELA)
Loading amount 30 µg
Specification HELA
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

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Verified customer

投稿 Nov 08 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - other (MEF)
Loading amount 30 µg
Specification MEF
Gel Running Conditions Reduced Denaturing
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 Nov 08 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cultured Cells (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton-X
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Username

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投稿 Oct 12 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cultured Cells (Embryonic Fibroblasts)
Specification Embryonic Fibroblasts
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton-x
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Username

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投稿 Oct 05 2010

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

Abcam user community

Verified customer

投稿 Nov 27 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (Brain cortex)
Loading amount 10 µg
Specification Brain cortex
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

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Verified customer

投稿 Oct 07 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - other (vero cells- mitochondrial fraction)
Loading amount 20 µg
Specification vero cells- mitochondrial fraction
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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投稿 Sep 29 2009

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