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Immunocytochemistry/ Immunofluorescence - Calnexin antibody (ab22595)
Immunocytochemistry/ Immunofluorescence - Calnexin antibody (ab22595)
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Western blot - Calnexin antibody (ab22595)
Western blot - Calnexin antibody (ab22595)
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Western blot - Calnexin - ER membrane marker antibody (ab22595)
Western blot - Calnexin - ER membrane marker antibody (ab22595)
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Product Name 

Calnexin - ER membrane marker antibody

 

See all Calnexin - ER membrane marker antibodies (10)...

Product type 

Primary antibodies

Description 

Rabbit polyclonal to Calnexin - ER membrane marker

Immunogen 

Synthetic peptide conjugated to KLH derived from within residues 550 to the C-terminus of Human Calnexin.

(Peptide available as ab23379.)

Reacts with 

Hu, Ms, Rat, Mmst

Predicted to react 
(species key)

Dog
(Due to sequence homology)

Tested applications 
(see key)

ICC/IF, IHC-P, WB


Abreviews 

Customer reviews
(see key)


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• ICC

1 Abreview

- average rating: 3 stars (out of 5)3 stars (out of 5)3 stars (out of 5)3 stars (out of 5)3 stars (out of 5)

• IHC-P

3 Abreviews

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• IHC-FoFr

1 Abreview

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• WB

4 Abreviews

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IHC-P
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IHC-P
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IHC-P
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WB
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Application notes 
(see key)

Recommended dilutions
ICC/IF: Use at a concentration of 5 µg/ml.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 90 kDa). Can be blocked with Calnexin peptide (ab23379).


Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.

Positive control 
(see definition)

ab22595 gave a positive result in the following Human Whole Cell Lysates:
Hela, MCF-7
Mouse Whole Cell Lystates:
NIH 3T3, MEF1
Mouse Tissue Lysates:
Brain, Liver, Heart, Kidney, Pancreas, Testis, Skeletal muscle, Spinal cord, Ovary
Rat Whole Cell Lysates:
PC12,
Rat Tissue Lysates:
Brain, Liver, Heart, Kidney

Cellular localization 

Endoplasmic reticulum

Research areas 

Neuroscience >> Sensory System >> Visual system
Tags & Cell Markers >> Subcellular Markers >> Organelles >> ER

Relevance 

Calnexin, also referred to as IP90, p88 and p90, is an ~90 kDa integral membrane protein of the endoplasmic reticulum (ER). Many resident ER proteins act as molecular chaperones and participate in the proper folding of polypeptides and their assembly into multisubunit proteins. Studies indicate that calnexin associates with the major histocompatability complex (MHC) class I heavy chains, partial complexes of the T cell receptor and B cell membrane immunoglobulin, but not with completed receptor complexes. It has been shown that calnexin is a chaperone that retains incompletely or improperly folded proteins in the ER. The sequence Lys-Asp-Glu-Leu (KDEL) or a closely related sequence, is present at the carboxy-terminus of soluble ER resident proteins such as GRP 78 and GRP 94 and protein disulfide isomerase. Integral membrane ER resident proteins, like calnexin, often lack this KDEL sequence but contain positively charged cytosolic residues that ensure ER retention. Calnexin contains a large ER luminal domain (461 amino acids), a transmembrane segment (22 amino acids), and a cytoplasmic tail (89 amino acids). These features distinguish calnexin from soluble ER chaperones that cannot interact with the transmembrane and cytosolic domains of integral membrane proteins. The amino acid sequence of calnexin is highly conserved among species, and shares regions of high sequence homology with calreticulin.

 

Alternative names Calnexin - ER membrane marker antibody (ab22595)

Database links 

The links below go to external sites and will open in a new browser window

Entrez Gene

    

403908    (Dog)
821    (Human)
12330    (Mouse)
29144    (Rat)

Omim

    

114217    (Human)

SwissProt

    

P24643    (Dog)
P27824    (Human)
P35564    (Mouse)
P35565    (Rat)

Raised in 

Rabbit

Clonality 

Polyclonal

Isotype 

IgG

Purity 

Immunogen affinity purified

Storage buffer 

Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4

Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS

Form 

Liquid

Concentration 

Batch dependent within range: 0.10 - 0.30 mg/ml

Concentration of lot no.     is  

Concentration not available for this lot.

Find concentration of your lot :

Storage instructions 

Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Store in the dark. Avoid repeated freeze / thaw cycles.

 

At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Calnexin - ER membrane marker antibody is on this datasheet. But please do contact us if you would like any reassurance!


See below for Calnexin - ER membrane marker antibody images, references, products related to ab22595 and other tools.

 

Calnexin - ER membrane marker antibody images:

Product image  Immunocytochemistry/ Immunofluorescence - Calnexin antibody (ab22595)

Immunocytochemistry/ Immunofluorescence - Calnexin antibody (ab22595)

ICC/IF image of ab22595 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab22595, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

 

Product image  Western blot - Calnexin antibody (ab22595)

Western blot - Calnexin antibody (ab22595)

All lanes : Calnexin - ER membrane marker antibody (ab22595) at 1 µg/ml

Lane 1 : HeLa (Human epithelial carcinoma cell line)
Lane 2 : U2OS Whole Cell Lysate
Lane 3 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 4 : HeLa (Human epithelial carcinoma cell line) with Calnexin peptide (ab23379) at 1 µg/ml
Lane 5 : U2OS Whole Cell Lysate with Calnexin peptide (ab23379) at 1 µg/ml
Lane 6 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate with Calnexin peptide (ab23379) at 1 µg/ml

Lysates/proteins at 20 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size : 90 kDa
Observed band size : 75 kDa (why is the actual band size different from the predicted?)


Recent batches of ab22595 (AP217379 and AP151845) detect a band of ~ 75 kDa in Hela, U2OS and MCF-7 lysates. This band is completely blocked by the immunizing peptide so we believe this represents Calnexin. Moreoever, a band of the same size is detected by other Calnexin antibodies tested.

 

Product image  Western blot - Calnexin - ER membrane marker antibody (ab22595)

Western blot - Calnexin - ER membrane marker antibody (ab22595)

All lanes : Calnexin - ER membrane marker antibody (ab22595) at 1/250 dilution

Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab46770)
Lane 3 : Brain (Mouse) Tissue Lysate (ab27253)
Lane 4 : Liver (Mouse) Tissue Lysate (ab7935)
Lane 5 : Heart (Mouse) Tissue Lysate (ab27255)
Lane 6 : Kidney (Mouse) Tissue Lysate (ab27254)
Lane 7 : Pancreas (Mouse) Tissue Lysate (ab29363)
Lane 8 : Testis (Mouse) Tissue Lysate - normal tissue (ab4027)
Lane 9 : Skeletal Muscle (Mouse) Tissue Lysate (ab29711)
Lane 10 : Spinal Cord (Mouse) Tissue Lysate (ab50253)
Lane 11 : Ovary (Mouse) Tissue Lysate (ab35808)
Lane 12 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate (ab50957)
Lane 13 : Brain (Rat) Tissue Lysate (ab7942)
Lane 14 : Liver (Rat) Tissue Lysate (ab27256)
Lane 15 : Heart (Rat) Tissue Lysate (ab7940)
Lane 16 : Kidney (Rat) Whole Cell Lysate - normal tissue (ab29480)

Lysates/proteins at 10 µg per lane.

Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size : 90 kDa
Observed band size : 80 kDa (why is the actual band size different from the predicted?)

 

Product image  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Calnexin - ER membrane marker antibody (ab22595)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Calnexin - ER membrane marker antibody (ab22595)

Kidney cortex using ab22595 shows clear cytoplasmic staining patterns. The visceral cells of the Glomerular tuft ( podocytes ) are strongly stained (indicated by red arrowheads). Distal convoluted tubular cells are generally moderately positive (with exceptions that are strongly positive). However, most of the cells that line the Proximal Convoluted Tubules (indicated by green arrowheads) are strongly positive.

Carl Hobbs, King`s College London, United Kingdom

See Abreview

 

References for Calnexin - ER membrane marker antibody (ab22595)

Specific publication references   This product has been used in: (two most recent references)

Wachten S et al. Distinct pools of cAMP centre on different isoforms of adenylyl cyclase in pituitary-derived GH3B6 cells. J Cell Sci 123:95-106 (2010). WB; Human. PubMed: 20016070

Takahashi Y et al. Identification of a novel palmitylation site essential for membrane association and isomerohydrolase activity of RPE65. J Biol Chem 284:3211-8 (2009). ICC/IF; Human. PubMed: 19049981
If you publish research using ab22595 please let us know so that we can cite the reference on this datasheet.

 

Search PubMed (MEDLINE) for references to Calnexin - ER membrane marker

 

Calnexin - ER membrane marker antibody - more information

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Calnexin - ER membrane marker antibody - related products:

Products for Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Slides

Products for Western blot

Loading controls

Lysates

 

Immunizing Peptide (Blocking) for Calnexin - ER membrane marker antibody

ab23379

  

Calnexin peptide

 

Immunizing Peptide for Calnexin - ER membrane marker antibody

 

Compatible Secondaries for Calnexin - ER membrane marker antibody

ab6564

  

Cy5 ® antibody

ab6717

  

FITC antibody

ab6720

  

Biotin antibody

ab6721

  

HRP antibody

ab6722

  

Alkaline Phosphatase antibody

ab6793

  

Texas Red ® antibody

ab7007

  

Phycoerythrin antibody

ab7019

  

Agarose antibody

ab27234

  

10nm Gold antibody

ab27235

  

5nm Gold antibody

ab27236

  

15nm Gold antibody

ab27237

  

20nm Gold antibody

ab27238

  

30nm Gold antibody

ab27239

  

10nm Gold antibody

ab27240

  

5nm Gold antibody

ab31193

  

Duck polyclonal to Rabbit IgG

ab60314

  

Chromeo™ 488 antibody

ab60315

  

Chromeo™ 494 antibody

ab60317

  

Chromeo™ 546 antibody

ab60319

  

Chromeo™ 642 antibody

 

Other Calnexin - ER membrane marker Antibodies

See all Calnexin - ER membrane marker antibodies (10)...

 

Calnexin - ER membrane marker antibody - other tools:

Contact Abcam with a Technical Enquiry about Calnexin - ER membrane marker antibody

 

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