bsm-54328R [Primary Antibody]
KDELR1 (3A6) Monoclonal Antibody
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DATASHEET

Host: Rabbit

Target Protein: KDELR1

Clonality: Monoclonal

Isotype: IgG

Entrez Gene: 10945

Swiss Prot: P24390

Source: Synthetic peptide corresponding to C-terminal human KDEL

Purification: Purified by Protein A.

Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

Storage: Store at 4°C for up to 2 weeks. For long term storage, store at -20°C in small aliquots to prevent freeze-thaw cycles.

Background:

Retention of resident soluble proteins in the lumen of the endoplasmic reticulum (ER) is achieved in both yeast and animal cells by their continual retrieval from the cis-Golgi, or a pre-Golgi compartment. Sorting of these proteins is dependent on a C-terminal tetrapeptide signal, usually lys-asp-glu-leu (KDEL) in animal cells, and his-asp-glu-leu (HDEL) in S. cerevisiae. This process is mediated by a receptor that recognizes, and binds the tetrapeptide-containing protein, and returns it to the ER. In yeast, the sorting receptor encoded by a single gene, ERD2, which is a seven-transmembrane protein. Unlike yeast, several human homologs of the ERD2 gene, constituting the KDEL receptor gene family, have been described. KDELR1 was the first member of the family to be identified, and it is structurally and functionally similar to the yeast ERD2. KDELR1 is required for the retention of luminal endoplasmic reticulum resident proteins via vesicular recycling. This receptor recognizes the C-terminal K-D-E-L motif. COPI-coated transport intermediates, either in the form of round vesicles or as tubular processes, mediate retrograde traffic of the KDEL receptor-ligand complexes. Also required for normal vesicular traffic through the Golgi.

Size: 100µL

Concentration: Lot Dependent

Applications: WB(1:300-5000)
FCM(1:20-100)
IHC-P(1:200-400)
IF(IHC-P)(1:50-200)
IF(ICC)(1:50-200)

Cross Reactive Species: Human
Mouse
Rat

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Lane 1: Rat testis tissue; Lane 2: Human placenta tissue; Lane 3: Mouse testis tissue; Lane 4: 293 Cells; Probed with KDELR1 (3A6) Monoclonal Antibody (bsm-54328R) at 1:1000 overnight at 4°C followed by a conjugated secondary antibody for 60 minutes at 37°C.


IF(ICC) staining with KDELR1 (3A6) Monoclonal Antibody (bsm-54328R) at 1:100 in A549 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.


IF(ICC) staining with KDELR1 (3A6) Monoclonal Antibody (bsm-54328R) at 1:200 in HepG2 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.


IF(ICC) staining with KDELR1 (3A6) Monoclonal Antibody (bsm-54328R) at 1:300 in 293T cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.


Paraformaldehyde-fixed and paraffin-embedded Human Placenta tissue incubated with KDELR1 (3A6) Monoclonal Antibody (bsm-54328R) at 1:200, overnight at 4°C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.


Paraformaldehyde-fixed and paraffin-embedded Human Stomach Cancer tissue incubated with KDELR1 (3A6) Monoclonal Antibody (bsm-54328R) at 1:200, overnight at 4°C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.


Paraformaldehyde-fixed and paraffin-embedded Mouse Small Intestine tissue incubated with KDELR1 (3A6) Monoclonal Antibody (bsm-54328R) at 1:200, overnight at 4°C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.


Paraformaldehyde-fixed and paraffin-embedded Rat Epididymis tissue incubated with KDELR1 (3A6) Monoclonal Antibody (bsm-54328R) at 1:100, overnight at 4°C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.


Flow cytometric analysis of HepG2 cells with KDELR1 (3A6) Monoclonal Antibody (bsm-54328R) at a 1:100 dilution (red) compared with an unlabeled control (cells without incubation with primary antibody; black).


4% Paraformaldehyde-fixed HeLa (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (KDEL) monoclonal Antibody, unconjugated (bsm-54328R) 1:100, 90 min at 37°C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37°C for 90 min . DAPI (blue, C02-04002) was used to stain the cell nuclei.


4% Paraformaldehyde-fixed HeLa (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (KDEL) monoclonal Antibody, unconjugated (bsm-54328R) 1:100, 90 min at 37°C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.


The HeLa (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.). Cells stained with Primary Antibody for 30 min at room temperature,followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-KDEL antibody (bsm-54328R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.


4% Paraformaldehyde-fixed HeLa (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (KDEL) monoclonal Antibody, unconjugated (bsm-54328R) 1:100, 90 min at 37°C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37°C for 90 min, then stained with Phalloidin/BF555 for 30 min at room temperature; DAPI (blue, C02-04002) was used to stain the cell nuclei.