PA5-27262

antibody from Invitrogen Antibodies

Targeting: SYK

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-27262

Provider

Invitrogen Antibodies

Product name

Syk Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: A431, Raji, mouse BMDM, mouse spleen. Predicted reactivity: Mouse (87%), Rat (87%), Pig (89%), Rhesus Monkey (98%), Bovine (80%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.32 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Blood flow guides sequential support of neutrophil arrest and diapedesis by PILR-β1 and PILR-α.

Li YT, Goswami D, Follmer M, Artz A, Pacheco-Blanco M, Vestweber D
eLife 2019 Aug 6;8

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of SYK in methanol-fixed A431 cells using a SYK polyclonal antibody (Product # PA5-27262) at a 1:200 dilution.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of SYK was performed using 70% confluent log phase Ramos cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with SYK Rabbit Polyclonal Antibody(Product # PA5-27262) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 3--figure supplement 1. PILRs modulate beta2 integrin activity via Syk. ( A ) PMN were pre-treated with non-crosslinking rabbit IgG or crosslinking polyclonal anti-PILRs for 15 min, and then stimulated with 100 ng/ml CXCL-1 for 0 s (unstimulated), 30 s, 60 s and 75 s, followed by fixation and cell lysis. Syk was immunoprecipitated and analyzed for phospho-Syk (4G10). ( B ) PMNs from WT (black), PILR-alpha -/- (red) or PILR-beta1 -/- (blue) were treated with or without 1 uM Syk-inhibitor PRT-060318 for 30 min. Cells were allowed to adhere to ICAM-1-Fc coated surfaces in the presence of 100 ng/ml CXCL-1 with or without PRT-060318. Adherent cells were counted. n >= 12 for each genotype from at least two experiments. Error bars, SEM. Groups were compared by 2-tailed t-test. ns, not significant, *p