Antibodies, Proteins, Kits and Reagents for Life Science

314 937 ₸ Product size

100 µl 314 937 ₸

Order now and get it on Tuesday March 02, 2021

Anti-CD8 beta antibody [EPR22331-54] (ab228965)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR22331-54] to CD8 beta
Suitable for: IP, IHC-P, WB, IHC-Fr
Reacts with: Mouse

Product name

Anti-CD8 beta antibody [EPR22331-54]
See all CD8 beta primary antibodies
Description

Rabbit monoclonal [EPR22331-54] to CD8 beta
Host species

Rabbit

Tested Applications & Species

Application	Species
IHC-Fr	Mouse
IHC-P	Mouse
IP	Mouse
WB	Mouse

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Mouse spleen, lymph node and thymus lysates. IP: Mouse thymus lysate. IHC-P: Mouse spleen and thymus tissue. IHC-Fr: Mouse thymus tissue.
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: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22331-54
Isotype

IgG
Research areas

Immunoprecipitation - Anti-CD8 beta antibody [EPR22331-54] (ab228965)

CD8 beta was immunoprecipitated from 0.35 mg of mouse thymus lysate with ab228965 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab228965 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

Lane 1: Mouse thymus lysate 10 μg (Input).
Lane 2: ab228965 IP in mouse thymus lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab22896 in mouse thymus lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
Immunohistochemistry (Frozen sections) - Anti-CD8 beta antibody [EPR22331-54] (ab228965)

Frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized mouse thymus tissue stained for CD8 beta (green) using ab228965 at 1/100 dilution in immunohistochemical analysis. The secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution. Specific membrane staining on T cells in the cortex of mouse thymus (PMID 12702501) is observed. The nuclear conter stain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 beta antibody [EPR22331-54] (ab228965)

Immunohistochemical analysis of paraffin-embedded mouse thymus tissue labeling CD8 beta with ab228965 at 1/500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on T cells in the cortex of mouse thymus (PMID: 12702501) is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.

The section was incubated with ab228965 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 beta antibody [EPR22331-54] (ab228965)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD8 beta with ab228965 at 1/500 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on T cells in periarterial lymphatic sheath of mouse spleen (PMID: 18776164) is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, the ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.

The section was incubated with ab228965 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.
Western blot - Anti-CD8 beta antibody [EPR22331-54] (ab228965)

All lanes : Anti-CD8 beta antibody [EPR22331-54] (ab228965) at 1/1000 dilution

Lane 1 : Mouse spleen lysate
Lane 2 : Mouse lymph node lysate
Lane 3 : Mouse thymus lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 24 kDa
Observed band size: 28-32 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

CD8 beta is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID: 11719190).

Exposure times:
Lanes 1-2: 37 seconds.
Lane 3: 6 seconds.
Anti-CD8 beta antibody [EPR22331-54] (ab228965)

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