Human/Mouse/Rat HSP70/HSPA1A Antibody

Detection of Human and Mouse HSP70/HSPA1A by Simple Western^TM.

Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line and C2C12 mouse myoblast cell line, loaded at 0.2 mg/mL. A specific band was detected for HSP70/HSPA1A at approximately 66 kDa (as indicated) using 0.5 µg/mL of Rabbit Anti-Human/Mouse/Rat HSP70/HSPA1A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1663). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Mouse HSP70/HSPA1A by Immunohistochemistry

RFA increases HSP70 levels and activates MyD88. a C57BL/6 mice were exposed to RF and skin HSP70, HSc70, and HSP90 levels were then analyzed by western blotting at 6 and 24 h (hr) using GAPDH as internal control. b Representative IHC images of HSP70 expression in RF-treated and non-treated skin at 24 h. Scale: 250 µm. c WT and MyD88 KO mice were exposed to RF or sham treatment followed by ID injection of 10 µg OVA into RF or sham-treated skin. Serum anti-OVA antibody titer was measured 2 weeks later. n = 4–5. d WT mice were intradermally injected with 100 µg Pepinh-Control or Pepinh-MyD, or the same volume of PBS 3 and 1 h before RF treatment and ID OVA immunization at 10 µg dose. OVA immunization alone served as control (No adjuvant). Serum anti-OVA antibody titer was measured 2 weeks later. n = 4–5. One-way ANOVA with Tukey’s multiple comparison test was used to compare differences between groups in c and d. *p < 0.05; **p < 0.01. NS not significant. Representative of two independent experiments in c and d Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30209303), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse HSP70/HSPA1A by Western Blot

RFA increases HSP70 levels and activates MyD88. a C57BL/6 mice were exposed to RF and skin HSP70, HSc70, and HSP90 levels were then analyzed by western blotting at 6 and 24 h (hr) using GAPDH as internal control. b Representative IHC images of HSP70 expression in RF-treated and non-treated skin at 24 h. Scale: 250 µm. c WT and MyD88 KO mice were exposed to RF or sham treatment followed by ID injection of 10 µg OVA into RF or sham-treated skin. Serum anti-OVA antibody titer was measured 2 weeks later. n = 4–5. d WT mice were intradermally injected with 100 µg Pepinh-Control or Pepinh-MyD, or the same volume of PBS 3 and 1 h before RF treatment and ID OVA immunization at 10 µg dose. OVA immunization alone served as control (No adjuvant). Serum anti-OVA antibody titer was measured 2 weeks later. n = 4–5. One-way ANOVA with Tukey’s multiple comparison test was used to compare differences between groups in c and d. *p < 0.05; **p < 0.01. NS not significant. Representative of two independent experiments in c and d Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30209303), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human HSP70/HSPA1A by Western Blot

EVs isolated from NHL and AIDS-NHL cell lines sequester rituximab and inhibit apoptosis in Ramos cells. (A) Characterization of extracellular vesicles isolated from NHL and AIDS-NHL cell lines. Western blots of EVs demonstrate the presence of classic exosome markers: HSP70, TSG101, CD81, CD9, and (B) CD63. 20 µg of EV protein lysate was loaded into each well. Imaging/exposure times of each blot: CD81, 60 s; HSP70, 75 s; CD9, 60 s; TSG101, 40 s; and CD63, 60 s. (C) CD20 concentrations measured in EVs by ELISA. Data is for one Luminex assay run. (D) Ramos cells were treated with rituximab (3 µg/ml) in the presence or absence of EVs isolated from NHL (Raji, Ramos) or AIDS-NHL cell lines (2F7, RRBL, R) cell lines, including OY6, a lymphoblastoid cell line of AIDS-NHL. 30 µg of total EV protein was used for each treatment. Shown are representative flow cytometry plots of Ramos cells double stained with Propidium iodide (PI) and Annexin V-FITC after treatment with rituximab in the presence of 2F7 EVs or Raji EVs to evaluate subpopulations of cells undergoing apoptosis. Percentage values are shown in each quadrant of each plot. (E) Quantitative data presented as bar graphs showing the fold increase of apoptosis: Ratio of % double positive cells (PI + and Annexin V+) after treatment with rituximab/untreated Ramos cells. Results are from two to four independent experiments. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40646161), licensed under a CC-BY license. Not internally tested by R&D Systems.