Antibodies, Proteins, Kits and Reagents for Life Science

291 484 ₸ Product size

100 µl 291 484 ₸

Order now and get it on Tuesday March 09, 2021

Anti-Hsp90 antibody [EPR16621-67] (ab203126)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR16621-67] to Hsp90
Suitable for: ICC/IF, IP, Flow Cyt, IHC-P, WB
Reacts with: Mouse, Rat, Human, Recombinant fragment

Product name

Anti-Hsp90 antibody [EPR16621-67]
See all Hsp90 primary antibodies
Description

Rabbit monoclonal [EPR16621-67] to Hsp90
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Mouse Human
IHC-P	Mouse Rat Human
IP	Human
WB	Mouse Rat Human Recombinant fragment

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Hsp90 alpha recombinant protein fragment (GST-tag): aa533-732; Hsp90 beta recombinant protein fragment (His-Tag®): aa525-724 ; Human fetal brain, fetal heart and fetal kidney lysates; Mouse and rat brain, heart and kidney lysates; HeLa, HEK-293, K562, Jurkat, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates. IHC-P: Human testis, Human lung cancer, mouse cerebral cortex and rat testis tissues. ICC/IF: HeLa and NIH/3T3 cells. Flow cytometry: Jurkat cells. IP: HeLa whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR16621-67
Isotype

IgG
Research areas

Western blot - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

All lanes : Anti-Hsp90 antibody [EPR16621-67] (ab203126) at 1/10000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse kidney lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat heart lysate
Lane 6 : Rat kidney lysate
Lane 7 : C6 (Rat glial tumor cells) whole cell lysate
Lane 8 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 10 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 85, 83 kDa
Observed band size: 90 kDa
why is the actual band size different from the predicted?

Exposure time: 3 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

Immunocytochemistry/Immunofluorescence analysis of 100% methanol-fixed NIH/3T3 (Mouse embryonic fibroblast) cells labeling Hsp90 alpha + beta with ab203126 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm staining on HeLa cell line. The nuclear counter stain is DAPI (blue).

Control - PBS instead of primary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Hsp90 alpha + beta with ab203126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm and weak nucleus staining on germ cells of Human testis is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

Immunocytochemistry/Immunofluorescence analysis of 100% methanol-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Hsp90 alpha + beta with ab203126 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm staining on HeLa cell line. The nuclear counter stain is DAPI (blue).

Control - PBS instead of primary antibody.
Flow Cytometry - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

Flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling Hsp90 alpha + beta with ab203126 at 1/350 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; 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.
Immunoprecipitation - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

Hsp90 alpha + beta was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab203126 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab203126 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: HeLa whole cell lysate 10 µg (Input).
Lane 2: ab203126 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab203126 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling Hsp90 alpha + beta with ab203126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm and weak nucleus staining on tumor cells of Human lung cancer is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

All lanes : Anti-Hsp90 antibody [EPR16621-67] (ab203126) at 1/10000 dilution

Lane 1 : Hsp90 alpha recombinant protein fragment (GST-tag): aa533-732
Lane 2 : Hsp90 beta recombinant protein fragment (His-Tag®): aa525-724

Lysates/proteins at 0.01 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 85, 83 kDa

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Human Hsp90 alpha and Hsp90 beta recombinant protein fragments were made in house.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling Hsp90 alpha + beta with ab203126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm and nucleus staining on neuron of mouse cerebral cortex is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

All lanes : Anti-Hsp90 antibody [EPR16621-67] (ab203126) at 1/10000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 85, 83 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling Hsp90 alpha + beta with ab203126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm and weak nucleus staining on germ cells of rat testis is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Hsp90 antibody [EPR16621-67] (ab203126)

Lanes 1 & 3-4 : Anti-Hsp90 antibody [EPR16621-67] (ab203126) at 1/100000 dilution
Lane 2 : Anti-Hsp90 antibody [EPR16621-67] (ab203126) at 1/10000 dilution

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 2 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
Lane 3 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 85, 83 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?

Exposure time: 3 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Anti-Hsp90 antibody [EPR16621-67] (ab203126)

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