Skip to main content

Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker is a rabbit recombinant monoclonal antibody that is used to detect Calnexin in Flow cytometry (Intra), IHC-P, Western blot. Suitable for Human samples.

- Endoplasmic reticulum (ER) membrane marker
- Recombinant format for unrivaled batch-batch consistency
- Specificity confirmed with CANX knockout cell line validation
- Cited in over 75 publications


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (AB133615), expandable thumbnail
  • Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (AB133615), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (AB133615), expandable thumbnail
  • Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (AB133615), expandable thumbnail
  • Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (AB133615), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWBFlow Cyt (Intra)
Human
Tested
Tested
Tested
Mouse
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Human
Dilution info
1/4000
Notes

For unpurified, use 1/100 - 1/250.

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000 - 1/5000
Notes

-

Not recommended
Not recommended

Species
Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/360
Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Not recommended
Not recommended

Species
Mouse
Dilution info
-
Notes

-

Associated Products

Select an associated product type

15 products for Alternative Product

Target data

Function

Calcium-binding protein that interacts with newly synthesized monoglucosylated glycoproteins in the endoplasmic reticulum. It may act in assisting protein assembly and/or in the retention within the ER of unassembled protein subunits. It seems to play a major role in the quality control apparatus of the ER by the retention of incorrectly folded proteins. Associated with partial T-cell antigen receptor complexes that escape the ER of immature thymocytes, it may function as a signaling complex regulating thymocyte maturation. Additionally it may play a role in receptor-mediated endocytosis at the synapse.

Alternative names

Calnexin, IP90, Major histocompatibility complex class I antigen-binding protein p88, p90, CANX

Recommended products

Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker is a rabbit recombinant monoclonal antibody that is used to detect Calnexin in Flow cytometry (Intra), IHC-P, Western blot. Suitable for Human samples.

- Endoplasmic reticulum (ER) membrane marker
- Recombinant format for unrivaled batch-batch consistency
- Specificity confirmed with CANX knockout cell line validation
- Cited in over 75 publications

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR3633(2)
Purification technique
Affinity purification Protein A
Specificity

Recognizes ER membrane, mitochondria and cis-Golgi.

Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Stable for 12 months at -20°C

Notes

Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), IHC-P, WB in human samples.

Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615) has been cited over 78 times in peer reviewed journals and is trusted by the scientific community.

Abcam's high quality manufacturing and validation processes ensure Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

The specificity of Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615) has been confirmed by testing in knockout samples.

Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615) specifically detects Calnexin (UniProt ID: P27824; Molecular weight: 65kDa) and is sold in 100 µL and 1 mL selling sizes.

Conjugation-ready, carrier free format available for antibody clone EPR3633(2) - Anti-Calnexin antibody [EPR3633(2)] - BSA and Azide free ab225542.

Antibody clone EPR3633(2) is also available pre-conjugated to a variety of labels for your convenience - HRP, Alexa Fluor® 594, APC, PE, Alexa Fluor® 488, Alexa Fluor® 647, Alexa Fluor® 568, Alexa Fluor® 555 (HRP Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker ab195198, Alexa Fluor® 594 Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker ab203439, APC Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker ab310822, PE Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker ab310899, Alexa Fluor® 488 Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker ab310982, Alexa Fluor® 647 Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker ab311104, Alexa Fluor® 568 Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker ab312999, Alexa Fluor® 555 Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker ab313204).

References regarding specificity:

Horner SM et al. Mitochondrial-associated endoplasmic reticulum membranes (MAM) form innate immune synapses and are targeted by hepatitis C virus. Proc Natl Acad Sci U S A 108:14590-5 (2011). PubMed: 21844353

Myhill N et al. The subcellular distribution of calnexin is mediated by PACS-2. Mol Biol Cell 19:2777-88 (2008). PubMed: 18417615

Yoshimura SI et al. Direct targeting of cis-Golgi matrix proteins to the Golgi apparatus. J Cell Sci 114:4105-15 (2001). PubMed: 11739642

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

Calnexin also known as Canx is a type I integral membrane protein of the endoplasmic reticulum (ER) involved in the process of protein folding. This chaperone protein has an approximate molecular weight of 90 kDa and is known for its role in the quality control of glycoproteins. Calnexin is expressed in the ER of cells where it interacts with nascent polypeptides to ensure proper folding and assembly contributing to cellular homeostasis. It exhibits its function through its lectin-like domain that binds to sugar moieties on glycoproteins.

Biological function summary

Calnexin facilitates the proper folding of newly synthesized proteins by forming a complex with another chaperone protein called ERp57. This interaction helps in creating the correct disulfide bonds in glycoproteins which is essential for their stability and functionality. The complex often referred to as the calnexin cycle is critical in preventing the aggregation and misfolding of proteins within the ER. This process ensures that only correctly folded proteins proceed to the Golgi apparatus for further processing and transport.

Pathways

Calnexin plays an important role in the ER-associated degradation (ERAD) pathway and the unfolded protein response (UPR). In these pathways calnexin ensures that misfolded proteins are retained in the ER or targeted for degradation preventing cellular stress. Calnexin is associated with proteins such as calreticulin another chaperone protein with a similar function in the ER. Together they maintain proteostasis within cells and protect against the accumulation of improperly folded proteins.

Associated diseases and disorders

Calnexin is linked to several conditions including cystic fibrosis and certain neurodegenerative diseases. In cystic fibrosis the misfolding and subsequent degradation of the CFTR protein are associated with calnexin's role in the ERAD pathway. Similarly in neurodegenerative diseases such as Alzheimer's disrupted protein folding and aggregation are linked to ER stress where calnexin and other chaperone proteins like BiP play a pivotal role in managing protein misfolding. Understanding calnexin's role in these disorders can contribute to developing strategies to mitigate faulty protein folding and its pathological consequences.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615)

    Immunohistochemical staining of paraffin embedded human pancreas with purified ab133615 at a working dilution of 1 in 4000. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP goat anti-rabbit (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615), expandable thumbnail

    Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615)

    Lanes 1- 2: Merged signal (red and green). Green - ab133615 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.

    ab133615 was shown to react with Calnexin in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line Human CANX (Calnexin) knockout HEK-293T cell line ab255368 (knockout cell lysate Human CANX (Calnexin) knockout HEK-293T cell lysate ab263805) was used. Wild-type HEK-293T and CANX knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133615 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615) at 1/1000 dilution

    Lane 1: Wild-type HEK-293T cell lysate at 20 µg

    Lane 2: CANX knockout HEK-293T cell lysate at 20 µg

    Lane 2: Western blot - Human CANX (Calnexin) knockout HEK-293T cell line (Human CANX (Calnexin) knockout HEK-293T cell line ab255368)

    Performed under reducing conditions.

    Predicted band size: 68 kDa

    Observed band size: 90 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615)

    Immunohistochemical staining of paraffin embedded rat cardiac muscle with purified ab133615 at a working dilution of 1 in 4000. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP goat anti-rabbit (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615), expandable thumbnail

    Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615)

    Lanes 1 - 4: Merged signal (red and green). Green - ab133615 observed at 80 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

    ab133615 was shown to recognize Calnexin when Calnexin knockout samples were used, along with additional cross-reactive bands. Wild-type and Calnexin knockout samples were subjected to SDS-PAGE. ab133615 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    All lanes: Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615) at 1/1000 dilution

    Lane 1: Wild-type HAP1 cell lysate at 20 µg

    Lane 2: Calnexin knockout HAP1 cell lysate at 20 µg

    Lane 3: THP1 cell lysate at 20 µg

    Lane 4: Raw264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cell lysate at 20 µg

    Predicted band size: 68 kDa

  • Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615), expandable thumbnail

    Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615)

    Lanes 1 - 4: Merged signal (red and green). Green - ab133615 observed at 80 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

    This western blot image is a comparison between ab133615 and a competitor's top cited rabbit polyclonal antibody.

    All lanes: Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615)

    Lane 1: Wild-type HAP1 cell lysate at 20 µg

    Lane 2: Calnexin knockout HAP1 cell lysate at 20 µg

    Lane 3: THP1 cell lysate at 20 µg

    Lane 4: Raw264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cell lysate at 20 µg

    Predicted band size: 68 kDa

  • Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615), expandable thumbnail

    Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

    All lanes: Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615) at 1/1000 dilution

    All lanes: HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate at 20 µg

    Secondary

    All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 68 kDa

    Observed band size: 90 kDa

  • Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615), expandable thumbnail

    Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

    All lanes: Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615) at 1/5000 dilution

    All lanes: HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg

    Secondary

    All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 68 kDa

    Observed band size: 90 kDa

  • Flow Cytometry (Intracellular) - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615)

    Overlay histogram showing HeLa cells stained with unpurified ab133615 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab133615, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Flow Cytometry (Intracellular) - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615)

    Overlay histogram showingHeLa cells fixed in 80%methanol and stained with purified ab133615 at a dilution of 1 in 360 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 150. Rabbit monoclonal IgG was used as an isotype control (black line) and cells without incubation with antibody were used as a negative control (blue line).

  • Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615), expandable thumbnail

    Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615)

    All lanes: Western blot - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615) at 1/1000 dilution

    Lane 1: HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate at 10 µg

    Lane 2: A431 (Human epidermoid carcinoma cell line) cell lysate at 10 µg

    Lane 3: SH-SY5Y (Human neuroblastoma cell line from bone marrow) cell lysate at 10 µg

    Lane 4: HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg

    Secondary

    All lanes: HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 68 kDa

    Observed band size: 90 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (ab133615)

    Immunohistochemical analysis of paraffin-embedded human kidney tissue labelled with unpurified ab133615 at 1/100 dilution.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com