Anti-Hsc70 antibody (ab112549)
Key features and details
- Rabbit polyclonal to Hsc70
- Suitable for: WB
- Knockout validated
- Reacts with: Human, Recombinant fragment
- Isotype: IgG
Overview
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Product name
Anti-Hsc70 antibody
See all Hsc70 primary antibodies -
Description
Rabbit polyclonal to Hsc70 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species WB HumanRecombinant fragment
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Recombinant Hsc70 protein; Human breast and kidney tissue lysates; and A431, HeLa, MCF7, MDA MB 231, and HEK293 whole cell lysates.
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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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 ab112549 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species WB HumanRecombinant fragmentAll applications MouseRatHorseChickenCowChinese hamsterOrangutanApplication Abreviews Notes WB Use a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 71 kDa).Notes WB
Use a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 71 kDa).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: 395853 Chicken
- Entrez Gene: 281831 Cow
- Entrez Gene: 3312 Human
- Entrez Gene: 15481 Mouse
- Entrez Gene: 24468 Rat
- Omim: 600816 Human
- SwissProt: O73885 Chicken
- SwissProt: P11142 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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Western blot - Anti-Hsc70 antibody (ab112549)All lanes : Anti-Hsc70 antibody (ab112549)
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: 70 kDa why is the actual band size different from the predicted?Lanes 1- 4: Merged signal (red and green). Green - ab112549 observed at 70 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab112549 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. ab112549 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 µg/ml 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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Western blot - Anti-Hsc70 antibody (ab112549)All lanes : Anti-Hsc70 antibody (ab112549) at 1 µg/ml
Lane 1 : Wild-type A431 whole cell lysate
Lane 2 : HSPA8 knockout A431 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : MCF7 whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 73 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab112549 observed at 73 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab112549 was shown to react with HSPA8 in A431 wild-type cells in Western blot. Loss of signal was observed when HSPA8 knockout sample was used. A431 wild-type and HSPA8 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab112549 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-Hsc70 antibody (ab112549)All lanes : Anti-Hsc70 antibody (ab112549) at 1 µg/ml
Lane 1 : Recombinant Protein:Hsc70 (Active)
Lane 2 : Human breast tissue lysate - total protein (ab30090)
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 4 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 5 : Human kidney tissue lysate - total protein (ab30203)
Lane 6 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 8 minutes
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab112549 has not yet been referenced specifically in any publications.
Images
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Western blot - Anti-Hsc70 antibody (ab112549)All lanes : Anti-Hsc70 antibody (ab112549)
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: 70 kDa why is the actual band size different from the predicted?Lanes 1- 4: Merged signal (red and green). Green - ab112549 observed at 70 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab112549 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. ab112549 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 µg/ml 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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Western blot - Anti-Hsc70 antibody (ab112549)All lanes : Anti-Hsc70 antibody (ab112549) at 1 µg/ml
Lane 1 : Wild-type A431 whole cell lysate
Lane 2 : HSPA8 knockout A431 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : MCF7 whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 73 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab112549 observed at 73 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab112549 was shown to react with HSPA8 in A431 wild-type cells in Western blot. Loss of signal was observed when HSPA8 knockout sample was used. A431 wild-type and HSPA8 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab112549 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
-
Western blot - Anti-Hsc70 antibody (ab112549)All lanes : Anti-Hsc70 antibody (ab112549) at 1 µg/ml
Lane 1 : Recombinant Protein:Hsc70 (Active)
Lane 2 : Human breast tissue lysate - total protein (ab30090)
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 4 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 5 : Human kidney tissue lysate - total protein (ab30203)
Lane 6 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 8 minutes
