Anti-CD47 antibody [EPR4150(2)] (ab108415)

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Anti-CD47 antibody [EPR4150(2)] (ab108415)

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商品描述

Product name

Anti-CD47 antibody [EPR4150(2)]
See all CD47 primary antibodies
Description

Rabbit monoclonal [EPR4150(2)] to CD47
Host species

Rabbit
Tested applications

Suitable for: WB,Flow Cyt,ICC/IF
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide within Human CD47 aa 250 to the C-terminus (intracellular). The exact sequence is proprietary.
Positive control
- Jurkat, HeLa, and K562 cell lysates; Jurkat cells
General notes

Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab^® patents.

We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

This product is a recombinant rabbit monoclonal antibody.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
Concentration
- 100 µl at 0.885 mg/ml
- Batch dependent within range: 40 µl at 0.809 - 0.885 mg/ml
- 10 µl at 0.885 mg/ml
Find concentration of your lot
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR4150(2)
Isotype

IgG

Flow Cytometry - Anti-CD47 antibody [EPR4150(2)] (ab108415)

Overlay histogram showing K562 cells fixed in 4% PFA and stained with purified ab108415 at a dilution of 1/180 (red line). The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit at a dilution of 1/500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
Immunocytochemistry/ Immunofluorescence - Anti-CD47 antibody [EPR4150(2)] (ab108415)

Immunofluorescence staining of Jurkat cells with purified ab108415 at a working dilution of 1/400, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab108415 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
Western blot - Anti-CD47 antibody [EPR4150(2)] (ab108415)

All lanes : Anti-CD47 antibody [EPR4150(2)] (ab108415) at 1/2000 dilution (purified)

Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate
Lane 3 : NIH/3T3 whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (HRP goat anti-rabbit IgG (H+L))

Predicted band size: 35 kDa
Observed band size: 52 kDa

why is the actual band size different from the predicted?
Western blot - Anti-CD47 antibody [EPR4150(2)] (ab108415)

All lanes : Anti-CD47 antibody [EPR4150(2)] (ab108415) at 1/2000 dilution (purified)

Lane 1 : Jurkat whole cell lysate
Lane 2 : HeLa whole cell lysate
Lane 3 : HEK293 whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (HRP goat anti-rabbit IgG (H+L))

Predicted band size: 35 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-CD47 antibody [EPR4150(2)] (ab108415)

All lanes : Anti-CD47 antibody [EPR4150(2)] (ab108415) at 1/1000 dilution (unpurified)

Lane 1 : Jurkat cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : K562 cell lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 35 kDa
Flow Cytometry - Anti-CD47 antibody [EPR4150(2)] (ab108415)

Flow cytometric analysis of CD47 in permeabilized Jurkat cells using unpurified ab108415 at 1/100 dilution (red) or a rabbit IgG (negative) (green)