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S-RMab® Hsp70 Recombinant Rabbit mAb (SDT-R016)

S-RMab® Hsp70 Recombinant Rabbit mAb (SDT-R016)

Catalog Number: S0B0007 Application: WB,IHC-P,ICC,FCM,IP Reactivity: Human Conjugation: Unconjugated Brand: Starter
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Product Details

Product Specification


Host Rabbit
Antigen Hsp70
Synonyms Heat shock 70 kDa protein
Immunogen Recombinant Protein
Location Cytoplasm, Nucleus
Accession P0DMV8,P0DMV9
Clone Number SDT-R016
Antibody Type Rabbit mAb
Application WB, IHC-P, ICC, ICFCM, IP
Reactivity Hu
Purification Protein A
Concentration 0.5mg/ml
Conjugation Unconjugated
Physical Appearance Liquid
Storage Buffer PBS, 40% Glycerol, 0.05%BSA, 0.03% Proclin 300
Stability & Storage 12 months from date of receipt / reconstitution, -20 °C as supplied.

Dilution


application dilution species
ICFCM 1:500 null
IHC-P 1:1000 null
IP 1:25 null
WB 1:2000-1:50000 null
ICC 1:100-1:500 null

Background

The 70 kilodalton heat shock proteins (Hsp70s or DnaK) are a family of conserved ubiquitously expressed heat shock proteins. Proteins with similar structure exist in virtually all living organisms. Intracellularly localized Hsp70s are an important part of the cell's machinery for protein folding, performing chaperoning functions, and helping to protect cells from the adverse effects of physiological stresses. Additionally, membrane-bound Hsp70s have been identified as a potential target for cancer therapies and their extracellularly localized counterparts have been identified as having both membrane-bound and membrane-free structures.

Picture

Western Blot

WB result of Hsp70 Rabbit mAb                

Primary antibody : Hsp70 Rabbit mAb at 1/1000 dilution
Lane 1: Hela whole cell lysate 20 µg
Lane 2: A431 whole cell lysate 20 µg
Lane 3: Jurkat whole cell lysate 20 µg
Lane 4: K562 whole cell lysate 20 µg


Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution


Predicted MW: 70 kDa
Observed MW: 70 kDa

FC

Flow cytometric analysis of HeLa cells labelling Hsp70 antibody at 1/500 (0.1ug) dilution/ (red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody. 

IP

Hsp70 Rabbit mAb at 1/25 dilution (2µg) immunoprecipitating Hsp70 in 0.4mg HeLa whole cell lysate.
Western blot was performed on the immunoprecipitate using Hsp70 Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/400 dilution.
Lane 1: HeLa whole cell lysate 10µg (input)
Lane 2: Hsp70 Rabbit mAb IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in HeLa whole cell lysate
Predicted MW: 70 kDa
Observed MW: 70 kDa

Immunohistochemistry

IHC shows positive staining in paraffin-embedded human testis.

Anti-Hsp70 antibody was used at 1/2000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP) ready to use. 

Counterstained with hematoxylin.

Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human prosate.

Anti-Hsp70 antibody was used at 1/2000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP) ready to use. 

Counterstained with hematoxylin.

Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human  lung adenocarcinoma.

Anti-Hsp70 antibody was used at 1/2000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP) ready to use. 

Counterstained with hematoxylin.

Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human breast cancer.

Anti-Hsp70 antibody was used at 1/2000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP) ready to use. 

Counterstained with hematoxylin.

Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

Immunocytochemistry

ICC shows positive staining in NIH/3T3 cells. Anti-Hsp70 antibody was used at 1/100 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue).Counterstain with tubulin (red).

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