- PA5-34772 - Invitrogen Antibodies HSPA1A antibody

Gao F, Fan H, Xue L, Shou Z, Zhu F, Yu T, Chu S, Wei C, Liu C, Zuo D, Zuo D
Disease markers 2022;2022:7115181

Jia J, Liu G, Zhong J, Yan R, Song X, Zheng K, Ren Z, He Z, Zhu Q
Frontiers in microbiology 2022;13:865644

Pan J, Ma N, Zhong J, Yu B, Wan J, Zhang W
Molecular therapy. Nucleic acids 2021 Dec 3;26:970-986

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of HSP70 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with HSP70 Polyclonal Antibody (Product # PA5-34772) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear and cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of HSPA1 was performed in HeLa cells fixed in ice-cold MeOH for 5 min. Green: HSPA1 Polyclonal Antibody (Product # PA5-34772) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry (Paraffin) analysis of HSPA1 was performed in paraffin-embedded rat brain tissue using HSPA1 Polyclonal Antibody (Product # PA5-34772) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: HSPA1A Polyclonal Antibody detects Hsp70 protein at cytosol on H1299 xenograft by immunohistochemical analysis. Sample: Paraffin-embedded H1299 xenograft. HSPA1A Polyclonal Antibody (Product # PA5-34772) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: HSPA1A Polyclonal Antibody immunoprecipitates Hsp70 protein in IP experiments. IP Sample: 1,000 µg HeLa whole cell lysate/extract A. 40 µg HeLa whole cell lysate/extract B. Control with 2.5 µg of preimmune rabbit IgG C. Immunoprecipitation of Hsp70 protein by 2.5 µg of HSPA1A Polyclonal Antibody (Product # PA5-34772) 12% SDS-PAGE The immunoprecipitated Hsp70 protein was detected by HSPA1A Polyclonal Antibody (Product # PA5-34772) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: HSPA1A Polyclonal Antibody immunoprecipitates Hsp70 protein in IP experiments. IP Sample: 1,000 µg HeLa whole cell lysate/extract A. 40 µg HeLa whole cell lysate/extract B. Control with 2.5 µg of preimmune rabbit IgG C. Immunoprecipitation of Hsp70 protein by 2.5 µg of HSPA1A Polyclonal Antibody (Product # PA5-34772) 12% SDS-PAGE The immunoprecipitated Hsp70 protein was detected by HSPA1A Polyclonal Antibody (Product # PA5-34772) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 8 Validation of transcriptome data by using qPCR and western blot (WB) (A and B) Expression analyses performed on selected genes yielded results superimposable with results obtained from RNA-seq analyses of (A) astrocytes and (B) microglia. Left: comparisons of DESeq2-analysis values between 12-month-olkd samples and 2-, 4-, 6-, and 9-month-old samples. Right: unpaired t tests for comparing two samples. (C) WB analysis of selected genes in astrocytic lysates. Loading was quantified by blotting with an anti-GAPDH antibody in cell lysates. (D) Quantification of the blot in (C) (unpaired t tests for comparing two samples). (E) WB analysis of selected genes in microglial lysates. Loading was quantified by blotting with an anti-GAPDH antibody in cell lysates. (F) Quantification of the blot in (E) (unpaired t tests for comparing two samples). Columns represent means +- SEM; ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 EV71 infection specially upregulates HSPA6 protein expression. (A) Western blotting analysis of HSPA1A, HSPA8, HSPB1, and viral VP1 expression in RD cells infected with EV71 for 8 or 24 h. (B) Western blotting analysis of HSPA6 expression in RD cells infected with EV71 for 8 or 24 h. The relative expression of HSPA6 was quantitatively analyzed based on the bands' intensities from the Western blotting analysis. PDL (10 muM) here was used as anti-EV71 control ( * p < 0.05, ** p < 0.01).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Expressions of HSPA1A and CHIP in colonic tissue. (a) The colonic protein levels of HSPA1A and CHIP measured by Western blotting. (b) Representative protein levels of HSPA1A/GAPDH. (c) Levels of HSPA1A mRNA in colonic tissue. (d) Representative protein levels of CHIP/GAPDH. (e) Levels of CHIP mRNA in colonic tissue. Data are shown as means +- SD ( n = 3). ** P < 0.01, *** P < 0.001, vs. DSS group.

PA5-34772