PRODUCT CODE: ET7107-86

KDEL Recombinant Rabbit Monoclonal Antibody [JB42-04] (ET7107-86)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of KDEL on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7107-86, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Rat testis tissue lysate<br />
Lane 2: Human placenta tissue lysate<br />
Lane 3: Mouse testis tissue lysate<br />
Lane 4: 293 cell lysate
  • Western blot analysis of KDEL on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7107-86, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Rat testis tissue lysate<br />
Lane 2: Human placenta tissue lysate<br />
Lane 3: Mouse testis tissue lysate<br />
Lane 4: 293 cell lysate
  • ICC staining of KDEL in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7107-86, 1/500) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of KDEL in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7107-86, 1/500) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of KDEL in 293T cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7107-86, 1/500) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-KDEL antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-86, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue using anti-KDEL antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-86, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue using anti-KDEL antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-86, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded rat epididymis tissue using anti-KDEL antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-86, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of KDEL was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7107-86, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of KDEL on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7107-86, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Rat testis tissue lysate
Lane 2: Human placenta tissue lysate
Lane 3: Mouse testis tissue lysate
Lane 4: 293 cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

KDEL Recombinant Rabbit Monoclonal Antibody [JB42-04] (ET7107-86)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

Rat testis tissue, Mouse testis tissue, 293, HepG2, A549, 293T, rat epididymis tissue, human placenta tissue, human stomach cancer tissue, mouse small intestine tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JB42-04

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

24/17 kDa(Predicted band size)

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500-1:2000

  • ICC

  • 1:400-1:800

  • IHC-P

  • 1:100-1:400

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

KDEL

SYNONYMS

ER lumen protein retaining receptor 1 antibody; ERD2.1 antibody; ERD21_HUMAN antibody; KDEL endoplasmic reticulum protein retention receptor 1 antibody; KDEL receptor 1 antibody; Kdelr1 antibody; Putative MAPK-activating protein PM23 antibody

SEQUENCE SIMILARITIES

Belongs to the ERD2 family.

POST-TRANSLATIONAL MODIFICATION

Phosphorylation by PKA at Ser-209 is required for endoplasmic reticulum retention function.

SUBCELLULAR LOCATION

Endoplasmic reticulum. Membrane.

FUNCTION

Soluble proteins in the endoplasmic reticulum (ER) contain a specific carboxy terminal sequence KDEL (Lys-Asp-Glu-Leu), and include the coat proteins required for vesicle budding from the ER, proteins that form retrograde vesicles on post-ER compartments, and integral membrane proteins that target vesicles to their correct destination. The retention of these soluble proteins in the ER depends on the interaction of the KDEL sequence with the corresponding KDEL receptor, also designated ERD2, in the Golgi apparatus. When KDEL proteins reach the Golgi complex, they are recognized by the KDEL receptor and transported retrograde in COPI-coated vesicles back to the ER. The small GTPase ADP-ribosylation factor 1 (ARF1), a regulator of vesicle transport, interacts with the KDEL receptor. Subsequently, this interaction allows the KDEL receptor to recruit a GTPase-activating protein (GAP) from the cytosol to membranes, which inactivates ARF1.