PA1-19378

antibody from Invitrogen Antibodies

Targeting: LCP2 SLP-76, SLP76

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-19378

Provider

Invitrogen Antibodies

Product name

SLP76 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Other

Description

This antibody reacts with SLP76, a 76kDa cytosolic adaptor protein that is involved in signaling of various hematopoietic cells, such as T cells, mast cells or neutrophils; in B cells, however, it is replaced by SLP65. Western Blot: Reducing conditions.

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

1 mg/mL

Storage

4° C, do not freeze

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-SLP76 Polyclonal Antibody (Product # PA1-19378) and a 70 kDa band corresponding to SLP76 was observed across the cell lines and tissue tested except in Raji and IMR-32. Whole cell extracts (30 µg lysate) of Jurkat (Lane 1), HEL 92.1.7 (Lane 2), THP-1 (Lane 3), Raji (Lane 4), IMR32 (Lane 5) and Mouse Thymus (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of SLP76 was performed using 70% confluent log phase Jurkat cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with SLP76 Polyclonal Antibody (PAb (412)) (Product # PA1-19378) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790) 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 Raji cells having no expression of SLP76. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry staining of human thymus (paraffin sections) with anti-SLP76 polyclonal antibody (Product # PA1-19378).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry staining of human thymus (paraffin sections) with anti-SLP76 Polyclonal antibody (Product # PA1-19378).