Antibodies, Proteins, Kits and Reagents for Life Science

291 484 ₸ Product size

100 µl 291 484 ₸

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Anti-SIRP alpha antibody [EPR16264] (ab191419)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR16264] to SIRP alpha
Suitable for: ICC/IF, WB, Flow Cyt
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-SIRP alpha antibody [EPR16264]
See all SIRP alpha primary antibodies
Description

Rabbit monoclonal [EPR16264] to SIRP alpha
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Rat
WB	Rat Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: THP1, SW480, C6, PC12, NIH 3T3 and Human fetal brain lysates. Flow Cyt: THP1 cells. ICC/IF: C6 cells
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: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR16264
Isotype

IgG
Research areas

Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)

All lanes : Anti-SIRP alpha antibody [EPR16264] (ab191419) at 1/1000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : SIRPA knockout HAP1 whole cell lysate
Lane 3 : THP1 whole cell lysate
Lane 4 : Jurkat whole cell lysate (negative control)

Lysates/proteins at 20 µg per lane.

Predicted band size: 55 kDa
Observed band size: 55 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab191419 observed at 55 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab191419 was shown to recognize SIRP alpha in wild-type HAP1 cells as signal was lost at the expected MW in SIRPA knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SIRPA knockout samples were subjected to SDS-PAGE. Ab191419 and ab9484 (Mouse anti-GAPDH 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.
Immunocytochemistry/ Immunofluorescence - Anti-SIRP alpha antibody [EPR16264] (ab191419)

Immunocytochemistry/ Immunofluorescence analysis of C6 (rat glial tumor glial cell) labeling SIRP alpha with ab191419 at 1/500. ab150077 Alexa Fluor^® 488 Goat anti-Rabbit at 1/1000 was used as the secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. DAPI was used to stain nuclei blue.

Confocal image showing membranous staining on C6 cell line.
Flow Cytometry - Anti-SIRP alpha antibody [EPR16264] (ab191419)

Flow Cytometric analysis of THP1 cells (2% paraformaldehyde-fixed) labeling SIRP alpha with ab191419 at 1/50 dilution (red) or a Rabbit monoclonal IgG (negative) (green), followed by Goat anti rabbit IgG (FITC) secondary at 1/150 dilution
Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)

All lanes : Anti-SIRP alpha antibody [EPR16264] (ab191419) at 1/5000 dilution

Lane 1 : THP1 cell lysate
Lane 2 : SW480 cell lysate
Lane 3 : Human fetal brain 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: 55 kDa
Western blot - Anti-SIRP alpha antibody [EPR16264] (ab191419)

All lanes : Anti-SIRP alpha antibody [EPR16264] (ab191419) at 1/1000 dilution

Lane 1 : C6 cell lysate
Lane 2 : PC12 cell lysate
Lane 3 : NIH 3T3 cell lysate

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: 55 kDa
Anti-SIRP alpha antibody [EPR16264] (ab191419)

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