Antibodies, Proteins, Kits and Reagents for Life Science

324 988 ₸ Product size

100 µl 324 988 ₸

Order now and get it on Tuesday March 02, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP1724Y] to Hsp27
Suitable for: WB, IHC-P
Knockout validated
Reacts with: Human

Product name

Anti-Hsp27 antibody [EP1724Y]
See all Hsp27 primary antibodies
Description

Rabbit monoclonal [EP1724Y] to Hsp27
Host species

Rabbit
Tested Applications & Species

Application Species
IHC-P
Human

WB
Human

See all applications and species data
Immunogen

Synthetic peptide within Human Hsp27 (N terminal). The exact sequence is proprietary.
Positive control
- WB: HeLa, HAP1 and MCF7 cell lysates. IHC-P: Human cervical carcinoma and human breast carcinoma tissues.
General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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.

Application	Species
IHC-P	Human
WB	Human

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer

pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
Concentration information loading...
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

EP1724Y
Isotype

IgG
Research areas

Western blot - Anti-Hsp27 antibody [EP1724Y] (ab62339)

All lanes : Anti-Hsp27 antibody [EP1724Y] (ab62339) at 1/500 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : HSPB1 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 23 kDa
Observed band size: 23 kDa

Lanes 1- 2: Merged signal (red and green). Green - ab62339 observed at 23 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab62339 was shown to react with Hsp27 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261738 (knockout cell lysate ab256945) was used. Wild-type HeLa and HSPB1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab62339 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp27 antibody [EP1724Y] (ab62339)

Immunohistochemical analysis of Hsp27 expression in paraffin embedded human cervical carcinoma (left) and human breast carcinoma (right) using 1/100 ab62339

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Western blot - Anti-Hsp27 antibody [EP1724Y] (ab62339)

All lanes : Anti-Hsp27 antibody [EP1724Y] (ab62339) at 1/500 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Hsp27 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : MCF7 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 23 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab62339 observed at 27 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab62339 was shown to specifically react with Hsp27 in wild-type HAP1 cells. No band was observed when Hsp27 knockout samples were used. Wild-type and Hsp27 knockout samples were subjected to SDS-PAGE. Ab62339 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-Hsp27 antibody [EP1724Y] (ab62339)

Anti-Hsp27 antibody [EP1724Y] (ab62339) at 1/500 dilution + HeLa cell lysate at 10 µg

Secondary
Goat anti-rabbit, HRP labeled at 1/2000 dilution

Predicted band size: 23 kDa
Observed band size: 27 kDa
why is the actual band size different from the predicted?
Anti-Hsp27 antibody [EP1724Y] (ab62339)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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