Anti-Hsp70 抗体 [EPR16892]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp70 antibody [EPR16892] (AB181606)

Immunohistochemical analysis of paraffin-embedded Human squamous cell carcinoma of cervix tissue labeling Hsp70 with ab181606 at 1/1000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasm staining on tumor cells of Human squamous cell carcinoma of cervix is observed. Counter stained with Hematoxylin.

Negative control : Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Hsp70 antibody [EPR16892] (AB181606)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling Hsp70with ab181606 at 1/230 dilution (red) compared with a rabbit monoclonal IgG control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Hsp70 antibody [EPR16892] (AB181606)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Hsp70 with ab181606 at 1/50 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Nuclear and cytoplasm staining on HeLa cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (Tubulin mouse mAb) at 1/500 and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

The negative controls are as follows;
1. ab181606 at 1/50 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab 150077 (Goat anti rabbit IgG (Alexa Fluor®488) secondary antibody at 1/400 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp70 antibody [EPR16892] (AB181606)

Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling Hsp70 with ab181606 at 1/1000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm and Nucleus staining on neuron cells of mouse cerebral cortex tissue is observed. Counter stained with Hematoxylin.

Negative control : Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp70 antibody [EPR16892] (AB181606)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Hsp70 with ab181606 at 1/1000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm staining on epithelial cells of mouse testis is observed. Counter stained with Hematoxylin.

Negative control : Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp70 antibody [EPR16892] (AB181606)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling Hsp70 with ab181606 at 1/1000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasm staining on epithelial cells of rat testis is observed. Counter stained with Hematoxylin.

Negative control : Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

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Western blot - Anti-Hsp70 antibody [EPR16892] (AB181606)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Hsp70 antibody [EPR16892] (ab181606) at 1/10000 dilution

Lane 1:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates at 20 µg

Lane 2:

293T (Human epithelial cells from embryonic kidney) whole cell lysates at 20 µg

Lane 3:

A431 (Human epidermoid carcinoma) whole cell lysates at 20 µg

Lane 4:

K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysates at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 70 kDa

Observed band size: 70 kDa

false

• WB

Supplier Data

Western blot - Anti-Hsp70 antibody [EPR16892] (AB181606)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Hsp70 antibody [EPR16892] (ab181606) at 1/1000 dilution

Lane 1:

Human fetal heart lysate at 10 µg

Lane 2:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 70 kDa

Observed band size: 70 kDa

false

• WB

Supplier Data

Western blot - Anti-Hsp70 antibody [EPR16892] (AB181606)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Hsp70 antibody [EPR16892] (ab181606) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse heart lysate at 10 µg

Lane 3:

Mouse kidney lysate at 10 µg

Lane 4:

Mouse spleen lysate at 10 µg

Lane 5:

Rat brain lysate at 10 µg

Lane 6:

Rat heart lysate at 10 µg

Lane 7:

Rat kidney lysate at 10 µg

Lane 8:

Rat spleen lysate at 10 µg

Lane 9:

C6 (Rat glial tumor cells) whole cell lysates at 10 µg

Lane 10:

RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates at 10 µg

Lane 11:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 10 µg

Lane 12:

NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 70 kDa

Observed band size: 70 kDa

false

• WB

CiteAb

Western blot - Anti-Hsp70 antibody [EPR16892] (AB181606)

Western Blotting using Anti-Hsp70 antibody [EPR16892], ab181606. Publication image from Qi, S. et al., 2020, Theranostics, 32042338. Legend direct from paper.

Immunological responses induced by LIT in the tumors, serum, and TDLNs. (A) Schematics of the procedures and timeline of GC + PTT treatment for CFP-B16 and analysis of LIT-induced antitumor immune response. (B) Proportions of neutrophils in immune cells in the treated primary tumors after various treatments at different times. (C) Proportions of neutrophils in immune cells with GC-RB (gated by CD45+ and RB+) in the tumors treated with GC-RB + PTT at different times. Data are presented as mean ± SD (n = 3-5 mice, two independent experiments). (D-F) Cytokine levels in serum (TNF-α, IL-6, and IL-1β) from mice at different times after various treatments. Data are presented as mean ± SD (n = 3-4 mice, two independent experiments). (G) HSP70 protein expression in the treated primary tumors 24 h after various treatments was analyzed using WB. (H) HSP70 and HMGB1 expressions in TDLNs at 48 h after different treatments were analyzed using WB (n = 3, two independent experiments). (I, J) The frequency of CD69+ in the CD4+(I) and CD8+(J) T cells of TDLNs 24 h after different treatments. (K) The frequency of mature DCs (CD11c+CD80+CD86+) in TDLNs 72 h after various treatments. Data are presented as mean ± SD (n = 4-7 mice, three independent experiments). Statistical analysis was performed using the unpaired t-test, and the one-way ANOVA test followed by the Bonferroni post-test. * P < 0.05, ** P < 0.01, *** P < 0.001, ns : not significant.

false

• WB

CiteAb

Western blot - Anti-Hsp70 antibody [EPR16892] (AB181606)

Western Blotting using Anti-Hsp70 antibody [EPR16892], ab181606. Publication image from Qi, S. et al., 2020, Theranostics, 32042338. Legend direct from paper.

Immunological responses induced by LIT in the tumors, serum, and TDLNs. (A) Schematics of the procedures and timeline of GC + PTT treatment for CFP-B16 and analysis of LIT-induced antitumor immune response. (B) Proportions of neutrophils in immune cells in the treated primary tumors after various treatments at different times. (C) Proportions of neutrophils in immune cells with GC-RB (gated by CD45+ and RB+) in the tumors treated with GC-RB + PTT at different times. Data are presented as mean ± SD (n = 3-5 mice, two independent experiments). (D-F) Cytokine levels in serum (TNF-α, IL-6, and IL-1β) from mice at different times after various treatments. Data are presented as mean ± SD (n = 3-4 mice, two independent experiments). (G) HSP70 protein expression in the treated primary tumors 24 h after various treatments was analyzed using WB. (H) HSP70 and HMGB1 expressions in TDLNs at 48 h after different treatments were analyzed using WB (n = 3, two independent experiments). (I, J) The frequency of CD69+ in the CD4+(I) and CD8+(J) T cells of TDLNs 24 h after different treatments. (K) The frequency of mature DCs (CD11c+CD80+CD86+) in TDLNs 72 h after various treatments. Data are presented as mean ± SD (n = 4-7 mice, three independent experiments). Statistical analysis was performed using the unpaired t-test, and the one-way ANOVA test followed by the Bonferroni post-test. * P < 0.05, ** P < 0.01, *** P < 0.001, ns : not significant.

false