Overview

  • Product name

    Anti-Hsp105/HSP110 antibody [EPR4576]
    See all Hsp105/HSP110 primary antibodies
  • Description

    Rabbit monoclonal [EPR4576] to Hsp105/HSP110
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Hsp105/HSP110 aa 50-150. The exact sequence is proprietary.

  • Positive control

    • WB: HepG2, MCF-7, HeLa, C6, PC-12 and NIH/3T3 cell lysates and human cerebellum tissue lysate. ICC/IF: HeLa cells. IHC-P: Human thyroid gland carcinoma and liver carcinoma tissues. IP: NIH/3T3 cell lysate.
  • General notes

    A trial size is available to purchase for this antibody.

     This product was previously labelled as Hsp105

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR4576
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab109624 in the following tested applications.

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

Application Abreviews Notes
WB 1/1000 - 1/10000. Predicted molecular weight: 97 kDa.
IP 1/10 - 1/100.
IHC-P 1/50 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF 1/100 - 1/250.

Target

  • Function

    Prevents the aggregation of denatured proteins in cells under severe stress, on which the ATP levels decrease markedly. Inhibits HSPA8/HSC70 ATPase and chaperone activities.
  • Tissue specificity

    Highly expressed in testis. Present at lower levels in most brain regions, except cerebellum. Overexpressed in cancer cells.
  • Sequence similarities

    Belongs to the heat shock protein 70 family.
  • Post-translational
    modifications

    Phosphorylation on Ser-509 may be important for regulation of the HSPA8/HSC70 chaperone activity.
  • Cellular localization

    Cytoplasm.
  • Information by UniProt
  • Database links

  • Alternative names

    • Antigen NY CO 25 antibody
    • Antigen NY-CO-25 antibody
    • DKFZp686M05240 antibody
    • Heat shock 105kD alpha antibody
    • Heat shock 105kD antibody
    • Heat shock 105kD beta antibody
    • Heat shock 105kDa protein 1 antibody
    • Heat shock 105kDa protein antibody
    • Heat shock 105kDa/110kDa protein 1 antibody
    • Heat shock 110 kDa protein antibody
    • Heat shock 110kDa protein antibody
    • Heat shock protein 105 kDa antibody
    • HS105_HUMAN antibody
    • HSP105 antibody
    • HSP105A antibody
    • HSP105B antibody
    • HSP110 antibody
    • HSPH 1 antibody
    • Hsph1 antibody
    • KIAA0201 antibody
    • NY CO 25 antibody
    see all

Images

  • All lanes : Anti-Hsp105/HSP110 antibody [EPR4576] (ab109624) at 1/5000 dilution (purified)

    Lane 1 : HeLa whole cell lysate
    Lane 2 : HepG2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 97 kDa
    Observed band size: 105 kDa
    why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.
  • All lanes : Anti-Hsp105/HSP110 antibody [EPR4576] (ab109624) at 1/5000 dilution (purified)

    Lane 1 : C6 whole cell lysate
    Lane 2 : PC-12 whole cell lysate
    Lane 3 : NIH/3T3 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 97 kDa
    Observed band size: 105 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver carcinoma tissue labelling Hsp105/HSP110 with purified ab109624 at a dilution of 1/500. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Hsp105/HSP110 with purified ab109624 at a dilution of 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/250) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

  • ab109624 (purified) at 1/40 immunoprecipitating Hsp105/HSP110 in NIH/3T3 whole cell lysate.

    Lane 1 (input): NIH/3T3 whole cell lysate (10µg)

    Lane 2 (+): ab109624 + NIH/3T3 whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109624 in NIH/3T3 whole cell lysate.

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-Hsp105/HSP110 antibody [EPR4576] (ab109624) at 1/1000 dilution (unpurified)

    Lane 1 : HeLa cell lysate
    Lane 2 : HepG2 cell lysate
    Lane 3 : Human cerebellum lysate
    Lane 4 : MCF-7 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size: 97 kDa

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Hsp105/HSP110 with unpurified ab109624 at a dilution of 1/100.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid gland carcinoma tissue labelling Hsp105/HSP110 with unpurified ab109624 at a dilution of 1/50.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

References

This product has been referenced in:

  • Deane CAS & Brown IR Differential Targeting of Hsp70 Heat Shock Proteins HSPA6 and HSPA1A with Components of a Protein Disaggregation/Refolding Machine in Differentiated Human Neuronal Cells following Thermal Stress. Front Neurosci 11:227 (2017). ICC/IF ; Human . Read more (PubMed: 28484369) »
  • Deane CA & Brown IR Components of a mammalian protein disaggregation/refolding machine are targeted to nuclear speckles following thermal stress in differentiated human neuronal cells. Cell Stress Chaperones 22:191-200 (2017). Read more (PubMed: 27966060) »
See all 3 Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK-293)
Gel Running Conditions
Reduced Denaturing (10% Acrylamide)
Loading amount
20 µg
Specification
HEK-293
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 20 2013

Application
Western blot
Sample
Human Cell lysate - whole cell (Human fibroblasts)
Gel Running Conditions
Reduced Denaturing (12.5%)
Loading amount
100000 cells
Specification
Human fibroblasts
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 16 2013

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Human fibroblasts)
Total protein in input
100000 cells
Immuno-precipitation step
Protein A/G
Specification
Human fibroblasts

Abcam user community

Verified customer

Submitted Jan 16 2013

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