重组Anti-Hsp105/HSP110抗体[EPR4576]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Hsp105/HSP110 antibody [EPR4576] (AB109624)

Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Hsp105/HSP110 with unpurified ab109624 at a dilution of 1/100.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp105/HSP110 antibody [EPR4576] (AB109624)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid gland carcinoma tissue labelling Hsp105/HSP110 with unpurified ab109624 at a dilution of 1/50.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp105/HSP110 antibody [EPR4576] (AB109624)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver carcinoma tissue labelling Hsp105/HSP110 with purified ab109624 at a dilution of 1/500. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051 a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Hsp105/HSP110 antibody [EPR4576] (AB109624)

Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Hsp105/HSP110 with purified ab109624 at a dilution of 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077 an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291 a mouse anti-tubulin (1/1000) and ab150120 an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1 : primary antibody (1/250) and secondary antibody ab150120 an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).

Control 2 : ab7291 (1/1000) and secondary antibody ab150077 an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).

• IP

Lab

Immunoprecipitation - Anti-Hsp105/HSP110 antibody [EPR4576] (AB109624)

ab109624 (purified) at 1/40 immunoprecipitating Hsp105/HSP110 in NIH/3T3 whole cell lysate.

Lane 1 (input) : NIH/3T3 whole cell lysate (10μg)

Lane 2 (+) : ab109624 + NIH/3T3 whole cell lysate.

Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab109624 in NIH/3T3 whole cell lysate.

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Immunoprecipitation - Anti-Hsp105/HSP110 antibody [EPR4576] (ab109624)

Predicted band size: 96 kDa

Observed band size: 105 kDa

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• WB

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Western blot - Anti-Hsp105/HSP110 antibody [EPR4576] (AB109624)

All lanes:

Western blot - Anti-Hsp105/HSP110 antibody [EPR4576] (ab109624) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

HepG2 cell lysate at 10 µg

Lane 3:

Human cerebellum lysate at 10 µg

Lane 4:

MCF-7 cell lysate at 10 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

Predicted band size: 96 kDa

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• WB

Lab

Western blot - Anti-Hsp105/HSP110 antibody [EPR4576] (AB109624)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Hsp105/HSP110 antibody [EPR4576] (ab109624) at 1/5000 dilution

Lane 1:

HeLa whole cell lysate at 20 µg

Lane 2:

HepG2 whole cell lysate at 20 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 83 kDa,96 kDa

Observed band size: 105 kDa

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• WB

Lab

Western blot - Anti-Hsp105/HSP110 antibody [EPR4576] (AB109624)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Hsp105/HSP110 antibody [EPR4576] (ab109624) at 1/5000 dilution

Lane 1:

C6 whole cell lysate at 20 µg

Lane 2:

PC-12 whole cell lysate at 20 µg

Lane 3:

NIH/3T3 whole cell lysate at 20 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 96 kDa

Observed band size: 105 kDa

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• WB

CiteAb

Western blot - Anti-Hsp105/HSP110 antibody [EPR4576] (AB109624)

Hsp105/HSP110 western blot using anti-Hsp105/HSP110 antibody [EPR4576] ab109624. Publication image and figure legend from Kim, M. J., Serwa, R. A., et al., 2023, Nat Commun, PubMed 37433777.

ab109624 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab109624 please see the product overview.

Small HSP and HSP110 families have a tendency to be increased under mitochondrial stress.a RNA-seq analysis of HSPs gene expression log2 fold changes (log2FC) in NDUFA11 KO and NDUFA13 KO compared to WT HEK293T cells (n = 4). Up- and down-regulated genes (q-value < 0.05) are shown in green and pink, respectively. The intensity of the color shades depends on the level of expression change. Gray indicates genes with not statistically significant expression changes. b mRNA expression patterns of selected transcripts validated by RT-qPCR. The mRNA levels are presented as fold changes relative to WT. Data shown are mean ± SD (n = 3 biological replicates with two technical replicates). p-value from an ordinary one-way ANOVA with Dunnett's multiple comparisons test using GraphPad Prism. c Western blot analysis of HSPs expression performed in whole cell lysates of NDUFA11 KO, NDUFA13 KO and WT HEK293T cells. ACTB was used as a loading control. Data shown are representative of three independent experiments. d Quantification of HSPs in western blot analysis normalized to ACTB using ImageJ. The protein levels are presented as fold changes relative to WT. Data shown are mean ± SD (n = 3). p-value from two-sided, unpaired t-test using GraphPad Prism. Source data are provided as a Source Data file.

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