Antibodies, Proteins, Kits and Reagents for Life Science

294 835 ₸ Product size

100 µl 294 835 ₸

Order now and get it on Tuesday March 09, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP15041] to CD58
Suitable for: WB
Knockout validated
Reacts with: Human

Product name

Anti-CD58 antibody [EP15041]
See all CD58 primary antibodies
Description

Rabbit monoclonal [EP15041] to CD58
Host species

Rabbit
Tested Applications & Species

Application Species
WB
Human

See all applications and species data
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa, Hap1, Daudi, Raji, Jurkat, TF-1, SW480 and K562 cell lysates.
General notes
Previously labelled as LFA3.
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
WB	Human

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, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EP15041
Isotype

IgG
Research areas

Western blot - Anti-CD58 antibody [EP15041] (ab196648)

All lanes : Anti-CD58 antibody [EP15041] (ab196648) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CD58 knockout HeLa cell lysate
Lane 3 : HAP1 cell lysate
Lane 4 : Daudi cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 28 kDa

Lanes 1-4: Merged signal (red and green). Green - ab196648 observed at 43 kDa. Red - loading control ab8245 observed at 37 kDa.

ab196648 Anti-CD58 antibody [EP15041] was shown to specifically react with CD58 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265947 (knockout cell lysate ab257880) was used. Wild-type and CD58 knockout samples were subjected to SDS-PAGE. ab196648 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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.
Western blot - Anti-CD58 antibody [EP15041] (ab196648)

All lanes : Anti-CD58 antibody [EP15041] (ab196648) at 1/1000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : CD58 knockout HAP1 whole cell lysate

Lysates/proteins at 40 µg per lane.

Predicted band size: 28 kDa
Observed band size: 43 kDa
why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab196648 observed at 43 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab196648 was shown to specifically react with CD58 in wild-type HAP1 cells as signal was lost in CD58 knockout cells. Wild-type and CD58 knockout samples were subjected to SDS-PAGE. Ab196648 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 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-CD58 antibody [EP15041] (ab196648)

All lanes : Anti-CD58 antibody [EP15041] (ab196648) at 1/5000 dilution

Lane 1 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 2 : 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), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 28 kDa
Observed band size: 65-70 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-CD58 antibody [EP15041] (ab196648)

All lanes : Anti-CD58 antibody [EP15041] (ab196648) at 1/1000 dilution

Lane 1 : TF-1 (Human bone marrow erythroleukemia cells) whole cell lysate
Lane 2 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 28 kDa
Observed band size: 65-70 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-CD58 antibody [EP15041] (ab196648)

All lanes : Anti-CD58 antibody [EP15041] (ab196648) at 1/2000 dilution

Lane 1 : SW480 (Human colorectal adenocarcinoma cell line) control
Lane 2 : SW480 (Human colorectal adenocarcinoma cell line) de-glycosylation

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 28 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.
Observed band size : 65-70 (27, 28) kDa.
Anti-CD58 antibody [EP15041] (ab196648)

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