HSP70 Rabbit Polyclonal Antibody

• Catalog Number: orb157591
• Category: Antibodies
• Description: HSP70 Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Gallus, Rabbit, Sheep
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human HSP70 (500-600/641aa)
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:100-500, Flow-Cyt=1ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 70 kDa
• Target: HSPA1A
• UniProt ID: P0DMV9
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: HSP70; HSP-70; HSP 70; Heat shock 70 kDa protein 1
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: 293T. Primary Antibody (green line): Rabbit Anti-HSP70 antibody (orb157591), Dilution: 2 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-FITC, Dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Blank control: A549. Primary Antibody (green line): Rabbit Anti-HSP70 antibody (orb157591), Dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF488, Dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Blank control: A549. Primary Antibody (green line): Rabbit Anti-HSP70 antibody (orb157591), Dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF488, Dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (Human breast carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (HSP70) Polyclonal Antibody, Unconjugated (orb157591) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Mouse testis), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (HSP70) Polyclonal Antibody, Unconjugated (orb157591) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Rat testis), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (HSP70) Polyclonal Antibody, Unconjugated (orb157591) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Lane 1: Testis (Mouse) Lysate at 40 ug, Lane 2: Epididymis (Mouse) Lysate at 40 ug, Lane 3: Seminal vesicle (Mouse) Lysate at 40 ug, Lane 4: Liver (Mouse) Lysate at 40 ug, Lane 5: NIH/3T3 (Mouse) Cell Lysate at 30 ug, Lane 6: Testis (Rat) Lysate at 40 ug, Lane 7: Spleen (Rat) Lysate at 40 ug, Lane 8: Hela (Human) Cell Lysate at 30 ug, Lane 9: Jurkat (Human) Cell Lysate at 30 ug, Lane 10: HepG2 (Human) Cell Lysate at 30 ug, Lane 11: A549 (Human) Cell Lysate at 30 ug, Lane 12: MCF-7 (Human) Cell Lysate at 30 ug, Primary: Anti-HSP70 (orb157591) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 70 kD, Observed band size: 68 kD.

Tissue/Cell: U-87MG cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (HSP70) polyclonal Antibody, Unconjugated (orb157591) 1:100, 90 minutes at 37°C, followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.