Antibodies, Proteins, Kits and Reagents for Life Science

Anti-hHR23b antibody [2857D7a] (ab70602)

Key features and details

Mouse monoclonal [2857D7a] to hHR23b
Suitable for: IHC-P, WB
Knockout validated
Reacts with: Human, Recombinant fragment
Isotype: IgG1

Product name

Anti-hHR23b antibody [2857D7a]
See all hHR23b primary antibodies
Description

Mouse monoclonal [2857D7a] to hHR23b
Host species

Mouse
Tested Applications & Species

Application Species
IHC-P
Human

WB
Human

Recombinant fragment

See all applications and species data
Immunogen

Recombinant fragment derived from internal sequence of human hHR23b.
Positive control
- WB: HeLa, MCF7 and SW620 cell lysates; Recombinant human hHR23b. IHC-P: Human cervival carcinoma tissue.

Application	Species
IHC-P	Human
WB	Human Recombinant fragment

Form

Liquid
Storage instructions

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

pH: 7.40
Preservative: 0.05% Sodium azide
Constituents: 1% BSA, PBS
Concentration information loading...
Purity

Protein G purified
Purification notes

ab70602 was purified using protein G column chromatography from culture supernatant of hybridoma cultured in a medium containing bovine IgG-depleted (approximately 95%) fetal bovine serum and filtered through a 0.22µm membrane.
Clonality

Monoclonal
Clone number

2857D7a
Isotype

IgG1
Research areas

Western blot - Anti-hHR23b antibody [2857D7a] (ab70602)

All lanes : Anti-hHR23b antibody [2857D7a] (ab70602) at 1/500 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : RAD23B knockout HeLa cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : SW620 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution

Predicted band size: 43 kDa
Observed band size: 50 kDa
why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab70602 observed at 50 kDa. Red - loading control ab181602 observed at 36 kDa.

ab70602 Anti-hHR23b antibody [2857D7a] was shown to specifically react with hHR23b in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264961 (knockout cell lysate ab258623) was used. Wild-type and hHR23b knockout samples were subjected to SDS-PAGE. ab70602 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated at room temperature for 2.5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hHR23b antibody [2857D7a] (ab70602)

IHC image of ab70602 staining in human normal cervical carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab70602, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Anti-hHR23b antibody [2857D7a] (ab70602)

Anti-hHR23b antibody [2857D7a] (ab70602) at 0.2 µg/ml + Whole cell lysate from HeLa cells at 25 µg

Predicted band size: 43 kDa
Western blot - Anti-hHR23b antibody [2857D7a] (ab70602)

Anti-hHR23b antibody [2857D7a] (ab70602) at 0.2 µg/ml + immunising recombinant protein

Predicted band size: 43 kDa
Observed band size: 44 kDa why is the actual band size different from the predicted?

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