Log In Sign up
The largest database of trusted experimental protocols

Mouse anti hsp70

Manufactured by Santa Cruz Biotechnology
Visit Supplier
Sourced in United States

Mouse anti-HSP70 is a laboratory reagent used to detect the presence and quantify the levels of the heat shock protein 70 (HSP70) in biological samples. HSP70 is a highly conserved molecular chaperone protein that plays a crucial role in cellular protein folding and stress response. This antibody can be utilized in various immunoassay techniques, such as Western blotting, immunohistochemistry, and flow cytometry, to study the expression and distribution of HSP70 in different cell types and tissues.

Automatically generated - may contain errors

Lab products found in correlation

Pierce ip lysis buffer, by Thermo Fisher Scientific (4 mentions) Anykd criterion tgx gels, by Bio-Rad (2 mentions) Ezview red anti flag m2 affinity gel, by Merck Group (2 mentions) Chemidoc mp imaging system, by Bio-Rad (2 mentions) Odyssey imaging system, by LI COR (2 mentions) Mouse anti flag m2, by Merck Group (2 mentions) Rabbit anti bach2, by Abcam (2 mentions) 3xflag peptide, by Merck Group (2 mentions) Irdye 800cw goat anti mouse, by LI COR (2 mentions) Micro bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (2 mentions) Nitrocellulose membrane, by Bio-Rad (2 mentions) Complete protease inhibitor cocktail, by Roche (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Enhanced chemi luminescence (ecl), by Cell Signaling Technology (1 mentions) Rabbit anti alix, by Abcam (1 mentions) Rabbit anti tsg101, by Abcam (1 mentions) Rabbit anti calnexin, by Cell Signaling Technology (1 mentions) Chemidoc xrs system, by Bio-Rad (1 mentions) Mouse anti gapdh, by Merck Group (1 mentions)

Close spelling variants of this product

HSP70 (101 citations) Anti-HSP70 (55 citations) Mouse Hsp70 (2 citations) Mouse anti-Hsp70 (SC-24; (2 citations)

10 protocols using mouse anti hsp70

1

Western Blot Analysis of Protein Targets

Check if the same lab product or an alternative is used in the 5 most similar protocols
Western blotting of proteins was performed as described in published protocols (18 (link)). The cell lysates or immunoprecipitated proteins were analyzed by SDS-PAGE. After completion of electrophoresis, the proteins were transferred to a polyvinylidene fluoride or polyvinylidene difluoride (PVDF) membrane, followed by blocking at 4°C for a minimum of 1 h. The blots were probed overnight at 4°C with the primary antibodies. The antibodies used were as follows: mouse anti-Ago2 (Abnova), 1:1,000; rabbit anti-DICER1 (Bethyl), 1:8,000; rat anti-HA (Roche), 1:1,000; horseradish peroxidase (HRP)-conjugated anti-β-Actin (Sigma), 1:10000; rabbit anti-TRBP2 (Cell Signalling), 1:1000; rabbit anti-Drosha (Bethyl), 1:8,000; rabbit anti-P-p38 (Cell Signaling), 1:1,000; rabbit anti-P-ERK1/2 (Cell Signaling), 1:1,000; mouse anti-HSP70 (Santa Cruz Biotechnology), 1:1,000; rabbit anti-MSK1 (Cell Signaling), 1:1,000; mouse anti-HSP70 (Santa Cruz Biotechnology), 1:1,000; rabbit anti-cleaved PARP (Cell Signaling), 1:1,000; rabbit anti-cleaved caspase 9 (Cell Signaling), 1:1,000; and rabbit anti-P-Akt (Ser-473) (Cell Signaling), 1:1,000. Visualization of all Western blots was performed using an UVP BioImager 600 system equipped with VisionWorks Life Science software (UVP) V6.80. ImageJ software was used for densitometric analysis of blots for the relative quantification of bands.
Chatterjee S., Mukherjee I., Bhattacharjee S., Bose M., Chakrabarti S, & Bhattacharyya S.N. (2022). Target-Dependent Coordinated Biogenesis of Secondary MicroRNAs by miR-146a Balances Macrophage Activation Processes. Molecular and Cellular Biology, 42(4), e00452-21.
+ Open protocol
+ Expand
2

Exosome Characterization by Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Exosome-specific markers were detected by western blot analysis (WB) [19 (link),40 (link)]. Thirty micrograms of exosomes were used to perform SDS-PAGE, followed by semidry transfer. The following primary antibodies were used (overnight incubation): rabbit anti-ALIX (Abcam), rabbit anti-Tsg101 (Abcam), rabbit anti-calnexin (Cell Signaling Technology) and mouse anti-Hsp70 (Santa Cruz Biotechnology). The following peroxidase-conjugated antibodies were used: anti-rabbit (Abcam) and anti-mouse (Merck). The membranes were incubated in ECL (Cell Signaling Technology). Proteins were visualized using a ChemiDocTM XRS + system (BioRad). Protein lysate of human adipose MSCs were used as a positive control.
Villatoro A.J., Martín-Astorga M.D., Alcoholado C., Sánchez-Martín M.D, & Becerra J. (2021). Proteomic Analysis of the Secretome and Exosomes of Feline Adipose-Derived Mesenchymal Stem Cells. Animals : an Open Access Journal from MDPI, 11(2), 295.
+ Open protocol
+ Expand
3

Western Blot Analysis of ORAI1, HSP70, and Cytoskeletal Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Lysates were prepared using RIPA lysis buffer (Sigma) supplemented with protease inhibitors (Thermofisher Scientific). 30 and 50 μg of protein lysate was used for experiments with HEK293 and MCF7 cells respectively. Protein was loaded into a 4–12% Bis-Tris gel (Life Technologies) and then transferred onto a PVDF membrane (Millipore). After blocking for 1 hr. at room temperature with Olympus Blocking Buffer (LI-COR), membranes were incubated for 2 hours at room temperature with primary antibody diluted in Olympus Blocking Buffer and 0.1% Tween. The dilutions of primary antibodies were: rabbit-anti-ORAI1 (Sigma) 1:1000, mouse-anti-HSP70 (Santa Cruz) 1:4000, mouse-anti-GAPDH (Millipore) 1:3000, and mouse-anti-α-tubulin (CST) 1:2000. Following four washes with 0.1% TBST for five min. each, membranes were placed in secondary antibody diluted in Olympus Blocking Buffer and 0.1% Tween for 1 hr at room temperature. The secondaries were 1:10,000 680RD conjugated anti-mouse (LI-COR) and 1:5000 800CW conjugated anti-rabbit (LI-COR). After four further washes with 0.1% TBST for five min. each, and images of blots were taken using Odyssey Clx imaging system. Densitometry was quantified using ImageStudio Lite.
Azimi I., Stevenson R.J., Zhang X., Meizoso-Huesca A., Xin P., Johnson M., Flanagan J.U., Chalmers S.B., Yoast R.E., Kapure J.S., Ross B.P., Vetter I., Ashton M.R., Launikonis B.S., Denny W.A., Trebak M, & Monteith G.R. (2020). A New Selective Pharmacological Enhancer of the Orai1 Ca2+ Channel Reveals Roles for Orai1 in Smooth and Skeletal Muscle Functions. ACS Pharmacology & Translational Science, 3(1), 135-147.
+ Open protocol
+ Expand
4

Protein Quality Control Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols
Digitonin was obtained from Merck (Darmstadt, Germany), lactacystin was purchased from Enzo Life Sciences (Farmingdale, NY, USA), and epoxomicin was purchased from UBPBio (Oxfordshire, UK). The mouse monoclonal anti-HA antibodies, anti-rabbit Alexa594 and anti-mouse Alexa488 were obtained from Thermo Fisher Scientific (Waltham, MA, USA). The mouse monoclonal anti-EDEM1, monoclonal anti-β-actin-peroxidase, rabbit anti-APP C-terminal, as well as the secondary anti-rabbit HRP and anti-mouse HRP antibodies were from Merck (Darmstadt, Germany). The mouse monoclonal anti-APP were obtained from OriGene (Rockville, MD, USA), the rabbit anti-PDI were from Cell Signaling (Danvers, MA, USA). The mouse anti-calreticulin was from Enzo Life Sciences (Farmingdale, NY, USA), whereas mouse monoclonal anti-calnexin was from BD Biosciences (Bedford, MA, USA). The mouse anti-HSP70 was obtained from Santa Cruz Biotechnology (Dallas, TX, USA).
Nowakowska-Gołacka J., Czapiewska J., Sominka H., Sowa-Rogozińska N, & Słomińska-Wojewódzka M. (2021). EDEM1 Regulates Amyloid Precursor Protein (APP) Metabolism and Amyloid-β Production. International Journal of Molecular Sciences, 23(1), 117.
+ Open protocol
+ Expand
5

Western Blot Analysis of Apoptotic Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols
We separated 35 μg of protein by SDS-PAGE. After transfer to a polyvinylidene fluoride membrane (Millipore, Billerica, MA, USA), membranes were blocked with 5% nonfat dry milk in 0.1% Tween-PBS (TBST) and incubated overnight at 4°C with primary antibody in 5% bovine serum albumin/TBST [mouse anti-p53 (Cell Signaling, Danvers, MA, USA), mouse anti-cytochrome c (BD Biosciences, Franklin Lakes, NJ, USA), rabbit anti-cleaved caspase 3 (Cell Signaling), mouse anti-HSP 70 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse anti-cytochrome c oxidoreductase (COX-IV) (Invitrogen), mouse anti-histone H1b (Abcam, Cambridge, MA, USA)] followed by horseradish peroxidase (HRP)-conjugated secondary antibodies (goat anti-mouse IgG or goat anti-rabbit immunoglobulin G (IgG; Jackson ImmunoResearch, West Grove, PA, USA) (in 5% nonfat dry milk/TBST). HRP-conjugated mouse anti-β-actin (Sigma-Aldrich) was used for normalization of cytosolic samples. Blots were revealed using enhanced chemiluminescence Amersham ECL Western Blotting Detection Reagent (GE Healthcare, Little Chalfont, Buckinghamshire, UK) and visualized, captured, and analyzed using a LAS system and Image Quant software (GE Healthcare).
Maj M.A., Ma J., Krukowski K.N., Kavelaars A, & Heijnen C.J. (2017). Inhibition of Mitochondrial p53 Accumulation by PFT-μ Prevents Cisplatin-Induced Peripheral Neuropathy. Frontiers in Molecular Neuroscience, 10, 108.
+ Open protocol
+ Expand
6

Western Blot and FLAG Immunoprecipitation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Clarified protein extracts were prepared by lysis of cell pellets in Pierce™ IP lysis buffer (ThermoScientific) containing 1x cOmplete Protease Inhibitor cocktail (Roche). Protein concentrations were quantified (Micro BCA protein assay kit (ThermoScientific) to ensure equal loading. Proteins were resolved by SDS-PAGE on Any kD™Criterion™ TGX™ gels (Bio-Rad) and electrotransferred onto nitrocellulose membranes (Bio-Rad). Immunoblotting was performed using rabbit anti-BACH2 (Abcam), mouse anti-FLAG® M2 (Sigma), mouse anti-Hsp70 (SantaCruz Biotechnology) and goat anti-mouse IRDye® 800CW (Li-Cor) following by scanning on an Odyssey imaging system (Li-Cor Biotechnology) or anti-HA-HRP for development using SuperSignal® West Pico Chemiluminescent Substrate (ThermoScientific) and imaging on a ChemiDocTM MP Imaging system (Bio-Rad). FLAG IP was carried out using EZview™ Red Anti-FLAG® M2 Affinity gel (Sigma) according to manufacturer’s instructions followed by elution using 3X FLAG® Peptide (Sigma).
Afzali B., Grönholm J., Vandrovcova J., O’Brien C., Sun H.W., Vanderleyden I., Davis F.P., Khoder A., Zhang Y., Hegazy A.N., Villarino A.V., Palmer I.W., Kaufman J., Watts N.R., Kazemian M., Kamenyeva O., Keith J., Sayed A., Kasperaviciute D., Mueller M., Hughes J.D., Fuss I.J., Sadiyah M.F., Montgomery-Recht K., McElwee J., Restifo N.P., Strober W., Linterman M.A., Wingfield P.T., Uhlig H.H., Roychoudhuri R., Aitman T.J., Kelleher P., Lenardo M.J., O’Shea J.J., Cooper N, & Laurence A.D. (2017). BACH2 immunodeficiency illustrates an association between super-enhancers and haploinsufficiency. Nature immunology, 18(7), 813-823.
+ Open protocol
+ Expand
7

Western Blot and FLAG Immunoprecipitation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Clarified protein extracts were prepared by lysis of cell pellets in Pierce™ IP lysis buffer (ThermoScientific) containing 1x cOmplete Protease Inhibitor cocktail (Roche). Protein concentrations were quantified (Micro BCA protein assay kit (ThermoScientific) to ensure equal loading. Proteins were resolved by SDS-PAGE on Any kD™Criterion™ TGX™ gels (Bio-Rad) and electrotransferred onto nitrocellulose membranes (Bio-Rad). Immunoblotting was performed using rabbit anti-BACH2 (Abcam), mouse anti-FLAG® M2 (Sigma), mouse anti-Hsp70 (SantaCruz Biotechnology) and goat anti-mouse IRDye® 800CW (Li-Cor) following by scanning on an Odyssey imaging system (Li-Cor Biotechnology) or anti-HA-HRP for development using SuperSignal® West Pico Chemiluminescent Substrate (ThermoScientific) and imaging on a ChemiDocTM MP Imaging system (Bio-Rad). FLAG IP was carried out using EZview™ Red Anti-FLAG® M2 Affinity gel (Sigma) according to manufacturer’s instructions followed by elution using 3X FLAG® Peptide (Sigma).
Afzali B., Grönholm J., Vandrovcova J., O’Brien C., Sun H.W., Vanderleyden I., Davis F.P., Khoder A., Zhang Y., Hegazy A.N., Villarino A.V., Palmer I.W., Kaufman J., Watts N.R., Kazemian M., Kamenyeva O., Keith J., Sayed A., Kasperaviciute D., Mueller M., Hughes J.D., Fuss I.J., Sadiyah M.F., Montgomery-Recht K., McElwee J., Restifo N.P., Strober W., Linterman M.A., Wingfield P.T., Uhlig H.H., Roychoudhuri R., Aitman T.J., Kelleher P., Lenardo M.J., O’Shea J.J., Cooper N, & Laurence A.D. (2017). BACH2 immunodeficiency illustrates an association between super-enhancers and haploinsufficiency. Nature immunology, 18(7), 813-823.
+ Open protocol
+ Expand
8

Protein Extraction and Immunoblotting Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were briefly washed twice, harvested in ice-cold PBS, and then collected by centrifugation at 2,500 × g at 4°C for 5 min. Cells were lysed in RIPA buffer (50 mM Tris pH 8.0, 150 mM NaCl, 5 mM EDTA, 10% NP-40, 0.5% sodium deoxycholate, and 1 mM phenylmethylsulfonyl fluoride) and incubated for 10 min on ice. After adding SDS loading dye, cell lysates were resuspended by a 26 gauge needle with a syringe. Protein samples were resolved by SDS-PAGE and transferred to the NC membrane (10600002; GE Healthcare, USA). The primary antibodies used in immunoblotting were rabbit anti-MRPL2 (HPA064814, 1:1,000; Atlas Antibodies, Sweden), mouse anti-HSP70 (sc-66048, 1:500; Santa Cruz Biotechnology), mouse anti-β tubulin (66240-1-Ig, 1:5,000; Proteintech, China), mouse anti-eIF2a (sc-133132, 1:500; Santa Cruz Biotechnology), and rabbit anti-phospho eIF2a (ab32157, 1:10,000; Abcam). Horseradish peroxidase-conjugated secondary antibodies (Jackson Immunoresearch Laboratories) were used for detecting primary antibodies and reacted with ECL solution (1705061; Bio-Rad, USA). Luminescence signals were detected by iBright FL1500 (Invitrogen).
Park D., Yu Y., Kim J.H., Lee J., Park J., Hong K., Seo J.K., Lim C, & Min K.T. (2023). Suboptimal Mitochondrial Activity Facilitates Nuclear Heat Shock Responses for Proteostasis and Genome Stability. Molecules and Cells, 46(6), 374-386.
+ Open protocol
+ Expand
9

Neural Progenitor Cell Differentiation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Dulbecco’s modified medium/Ham’s F12 (DMEM/F12 1:1), Dulbecco’s Modified Eagle’s Medium (DMEM), B27 and anti-rabbit Alexa Fluor® 488-labelled were purchased from Life Technologies Corporation (Carlsbad, CA, Unites States). Foetal bovine serum (FBS) from Internegocios (Buenos Aires, Argentina) and (DCFH-DA) from Molecular Probes. Hydrogen Peroxide was purchased from Cicarelli (Santa Fe, Argentina). Rabbit anti-βIII-tubulin antibody, protease inhibitor cocktail, poly-D-lysine (PDL), epidermal growth factor (EGF) and human basic fibroblast growth factor (bFGF) were purchased from Sigma (St. Louis, MO, United States). Mousse anti-synaptophysin, mousse anti-β-Actin, mouse anti-GFAP, mouse anti-ALIX, mouse anti-TSG101 and mouse anti-HSP70 from Santa Cruz Biotechnology (Dallas, Texas, United States); rabbit anti-PSD-95 and Vybrant™ DiI Cell-Labelling Solution from Invitrogen. Rabbit anti-Olig2 and mouse anti-Cy3-labelled from Millipore (Massachusetts, United States).
Ocaña S.D., Magaquian D, & Banchio C. (2023). Neural stem cell-derived extracellular vesicles favour neuronal differentiation and plasticity under stress conditions. Frontiers in Molecular Neuroscience, 16, 1146592.
+ Open protocol
+ Expand
10

Exosome Protein Extraction and Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Proteins were extracted from exosomes and cells using RIPA lysis buffer with 100× protease inhibitor and 50× phosphatase inhibitor added. Total protein concentration was quantified using Pierce BCA Protein Assay Kit (catalog no. 23225, Thermo Fisher Scientific). Protein sample was mixed with sample buffer and loaded into a 10% sodium dodecyl-sulfate polyacrylamide (SDS-PAGE) gel to perform gel electrophoresis. Western blotting was performed using a protocol previously described in our research [51] . The primary antibodies used were mouse anti-CD9 (catalog no. sc-59140, Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-CD63 (catalog no. 10628D, Invitrogen, Thermo Fisher Scientific), mouse anti-CD81 (catalog no. sc-7637, Santa Cruz Biotechnology), mouse anti-HSP70 (catalog no. sc-32239, Santa Cruz Biotechnology), and rabbit anti-β-actin (catalog no. GTX109639, GeneTex, Hsinchu, Taiwan) using 1:500~1:1000 dilution. After incubation with appropriate secondary antibody at 1:5000 dilution, target proteins were detected using a T-Pro LumiFast Plus Chemiluminescence Detection Kit (catalog no. JT96-K002M, T-Pro Biotechnology, New Taipei City, Taiwan), and signals were visualized using Amersham Imager 600 (catalog no. 29083461, Cytiva Life Sciences).
Kurniawati I., Liu M.C., Hsieh C.L., Do A.D, & Sung S.Y. (2022). Targeting Castration-Resistant Prostate Cancer Using Mesenchymal Stem Cell Exosomes for Therapeutic MicroRNA-let-7c Delivery. Frontiers in bioscience (Landmark edition), 27(9).
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!