Recombinant Anti-Hsc70 antibody [EP1531Y] (ab51052)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1531Y] to Hsc70
- Suitable for: IP, Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Hsc70 antibody [EP1531Y]
See all Hsc70 primary antibodies -
Description
Rabbit monoclonal [EP1531Y] to Hsc70 -
Host species
Rabbit -
Tested applications
Suitable for: IP, Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human Hsc70 aa 600 to the C-terminus (C terminal). The exact sequence is proprietary.
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Positive control
- WB: A431, MCF7, and HeLa cell lysates. IHC-P: Human cerebral cortex tissue. ICC/IF: MCF7 cells. Flow Cyt (intra): MCF7 cells. IP: HeLa cell lysate.
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General notes
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1531Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab51052 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 |
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IP | (1) |
1/10 - 1/20.
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Flow Cyt (Intra) |
1/30 - 1/70.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/500 - 1/5000. Detects a band of approximately 71 kDa (predicted molecular weight: 71 kDa).
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IHC-P |
1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/100 - 1/250.
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Notes |
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IP
1/10 - 1/20. |
Flow Cyt (Intra)
1/30 - 1/70. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/500 - 1/5000. Detects a band of approximately 71 kDa (predicted molecular weight: 71 kDa). |
IHC-P
1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/250. |
Target
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Function
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. -
Tissue specificity
Ubiquitous. -
Sequence similarities
Belongs to the heat shock protein 70 family. -
Domain
The N-terminal 1-386 residues constitute the ATPase domain, while residues 387-646 form the peptide-binding domain. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR.
ISGylated. -
Cellular localization
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
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Database links
- Entrez Gene: 3312 Human
- Entrez Gene: 15481 Mouse
- Entrez Gene: 24468 Rat
- Omim: 600816 Human
- SwissProt: P11142 Human
- SwissProt: P63017 Mouse
- SwissProt: P63018 Rat
- Unigene: 180414 Human
see all -
Alternative names
- 2410008N15Rik antibody
- Constitutive heat shock protein 70 antibody
- Epididymis luminal protein 33 antibody
see all
Images
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All lanes : Anti-Hsc70 antibody [EP1531Y] (ab51052) at 1/500 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : HSPA8 knockout HeLa cell lysate
Lane 3 : A431 cell lysate
Lane 4 : MCF7 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 71 kDaLanes 1- 4: Merged signal (red and green). Green - ab51052 observed at 71 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab51052 was shown to react with Hsc70 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265664 (knockout cell lysate ab256944) was used. Wild-type HeLa and HSPA8 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab51052 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunofluorescence staining of MCF7 cells with purified ab51052 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab51052 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
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Immunohistochemical staining of paraffin embedded human cerebral cortex with purified ab51052 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Overlay histogram showing MCF7 cells fixed in 4% PFA and stained with purified ab51052 at a dilution of 1 in 70 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
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ab51052 (purified) at 1/20 immunoprecipitating Hsc70 in 10 μg HeLa (Lanes 1 and 2, observed at 71 kDa). Lane 3 - PBS. For western blotting, HRP Veriblot for IP Detection Reagent (ab131366) was used for detection (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
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All lanes : Anti-Hsc70 antibody [EP1531Y] (ab51052) at 1/5000 dilution (purified)
Lane 1 : C6 cell lysate
Lane 2 : PC-12 cell lysate
Lane 3 : Mouse heart tissue lysate
Lane 4 : Rat heart tissue lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution
Predicted band size: 71 kDa
Observed band size: 71 kDaBlocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Hsc70 antibody [EP1531Y] (ab51052) at 1/5000 dilution (purified)
Lane 1 : human fetal brain lysate
Lane 2 : HeLa cell lysate
Lane 3 : HEK293 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution
Predicted band size: 71 kDa
Observed band size: 71 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
References (41)
ab51052 has been referenced in 41 publications.
- Yi C et al. Differential Size Distribution and Estrogen Receptor Cargo of Oviductal Extracellular Vesicles at Various Stages of Estrous Cycle in Mice. Reprod Sci 29:2847-2858 (2022). PubMed: 35137347
- Li L et al. NVP-AUY922 alleviates radiation-induced lung injury via inhibition of autophagy-dependent ferroptosis. Cell Death Discov 8:86 (2022). PubMed: 35220409
- Galvão I et al. The Therapeutic Effect of Phosphopeptide P140 Attenuates Inflammation Induced by Uric Acid Crystals in Gout Arthritis Mouse Model. Cells 11:N/A (2022). PubMed: 36496970
- Kuijpers M et al. Neuronal Autophagy Regulates Presynaptic Neurotransmission by Controlling the Axonal Endoplasmic Reticulum. Neuron 109:299-313.e9 (2021). PubMed: 33157003
- Yang HC & Rhee WJ Single Step In Situ Detection of Surface Protein and MicroRNA in Clustered Extracellular Vesicles Using Flow Cytometry. J Clin Med 10:N/A (2021). PubMed: 33477255