Claudin 4 Recombinant Rabbit Monoclonal Antibody [PSH23-48]
cat.: HA724379
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC(Intra), IP
Clonality: Monoclonal
Clone number: PSH23-48
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer: 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 22 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human Claudin 4 aa 160-209.
Positive control: HT-29 (Human colorectal adenocarcinoma cells) cell lysate, Caco-2 (Human colorectal adenocarcinoma cells) cell lysate, MCF7 (Human breast cancer cells) cell lysate, SW480 (Human colorectal zdenocarcinoma cells) cell lysate, 4T1 (Mouse breast cancer cells) cell lysate, Mouse colon tissue lysate, Rat colon tissue lysate, Rat placenta tissue lysate.
Subcellular location: Cell junction, tight junction, Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC(Intra)
  IP
1:5,000-1:50,000
1:100
1:200
1:1,000
1-2μg/sample
Uniprot #: SwissProt: O14493 Human | O35054 Mouse
Entrez Gene: 304407 Rat
Alternative names: Claudin-4 Claudin4 CLD4_HUMAN CLDN 4 CLDN4 Clostridium perfringens enterotoxin receptor 1 Clostridium perfringens enterotoxin receptor CPE R CPE receptor CPE-R CPE-receptor CPER CPETR 1 CPETR CPETR1 hCPE R WBSCR8 Williams Beuren syndrome chromosome region 8 protein Williams-Beuren syndrome chromosomal region 8 protein
Images
HA724379_1.jpg Fig1: Western blot analysis of Claudin 4 on different lysates with Rabbit anti-Claudin 4 antibody (HA724379) at 1/50,000 dilution.

Lane 1: HT-29 (Human colorectal adenocarcinoma cells) cell lysate
Lane 2: SK-MEL-28 (Human malignant melanoma cells) cell lysate
Lane 3: Caco-2 (Human colorectal adenocarcinoma cells) cell lysate
Lane 4: MCF7 (Human breast cancer cells) cell lysate
Lane 5: SW480 (Human colorectal zdenocarcinoma cells) cell lysate
Lane 6: 4T1 (Mouse breast cancer cells) cell lysate

Lysates/proteins at 15 µg/Lane.
Exposure time: 6 seconds; ECL: K1801

Negative expression of claudin-4 protein in SK-MEL-28 cells is consistent with the predicted expression pattern.

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724379, 1/50,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 22 kDa
Observed band size: 22 kDa
HA724379_2.jpg Fig2: Western blot analysis of Claudin 4 on different lysates with Rabbit anti-Claudin 4 antibody (HA724379) at 1/5,000 dilution.

Lane 1: Mouse colon tissue lysate
Lane 2: Rat colon tissue lysate
Lane 3: Rat placenta tissue lysate

Lysates/proteins at 40 µg/Lane.
Exposure time: 10 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724379, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 22 kDa
Observed band size: 22 kDa
HA724379_3.jpg Fig3: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: HT-29 (Human colorectal adenocarcinoma cells)/SK-MEL-28 (Human malignant melanoma cells)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.

Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA724379, 1/100, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.

Counterstain: Beta tubulin (HA601187, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).

Negative expression of claudin-4 protein in SK-MEL-28 cells is consistent with the predicted expression pattern.
HA724379_4.jpg Fig4: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Colon
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724379, 1/200, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724379_5.jpg Fig5: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Endometrium
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724379, 1/200, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724379_6.jpg Fig6: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Liver
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724379, 1/200, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724379_7.jpg Fig7: Application: Flow Cytometry (Intra)

Species: Human
Sample: HT-29 (Human colorectal adenocarcinoma cells)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature.

Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA724379 (1/1,000) (Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
HA724379_8.jpg Fig8: Application: Flow Cytometry (Intra)

Species: Mouse
Sample: 4T1 (Mouse breast cancer cells)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature.

Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA724379 (1/1,000) (Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
HA724379_9.jpg Fig9: Immunoprecipitation (IP)

Claudin 4 was immunoprecipitated in 0.2 mg HT-29 (Human colorectal adenocarcinoma cells) cell lysate with HA724379 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724379 at 1/20,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HT-29 cell lysate (input)
Lane 2: HA724379 IP in HT-29 cell lysate
Lane 3: Rabbit IgG instead of HA724379 in HT-29 cell lysate

Exposure time: 10 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA724379, 1/20,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 22 kDa
Observed band size: 22 kDa
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.